WO2002064551A1 - Hiv protease inhibitors based on amino acid derivatives - Google Patents

Hiv protease inhibitors based on amino acid derivatives Download PDF

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WO2002064551A1
WO2002064551A1 PCT/CA2002/000190 CA0200190W WO02064551A1 WO 2002064551 A1 WO2002064551 A1 WO 2002064551A1 CA 0200190 W CA0200190 W CA 0200190W WO 02064551 A1 WO02064551 A1 WO 02064551A1
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group
compound
formula
carbon atoms
methylbenzenesulfonyl
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PCT/CA2002/000190
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French (fr)
Inventor
Brent Richard Stranix
Gilles Sauve
Abderrahim Bouzide
Alexandre Cote
Gervais Berube
Patrick Soucy
Yongsen Zhao
Jocelyn Yelle
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Pharmacor Inc.
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Priority to EP02704504A priority Critical patent/EP1377542B1/en
Priority to DE60221508T priority patent/DE60221508T2/en
Priority to CA002440931A priority patent/CA2440931C/en
Publication of WO2002064551A1 publication Critical patent/WO2002064551A1/en

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    • C07D295/00—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms
    • C07D295/16—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms acylated on ring nitrogen atoms
    • C07D295/18—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms acylated on ring nitrogen atoms by radicals derived from carboxylic acids, or sulfur or nitrogen analogues thereof
    • C07D295/182—Radicals derived from carboxylic acids
    • C07D295/185—Radicals derived from carboxylic acids from aliphatic carboxylic acids
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    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A61P31/14—Antivirals for RNA viruses
    • A61P31/18—Antivirals for RNA viruses for HIV
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
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    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C311/00—Amides of sulfonic acids, i.e. compounds having singly-bound oxygen atoms of sulfo groups replaced by nitrogen atoms, not being part of nitro or nitroso groups
    • C07C311/15—Sulfonamides having sulfur atoms of sulfonamide groups bound to carbon atoms of six-membered aromatic rings
    • C07C311/16—Sulfonamides having sulfur atoms of sulfonamide groups bound to carbon atoms of six-membered aromatic rings having the nitrogen atom of at least one of the sulfonamide groups bound to hydrogen atoms or to an acyclic carbon atom
    • C07C311/19—Sulfonamides having sulfur atoms of sulfonamide groups bound to carbon atoms of six-membered aromatic rings having the nitrogen atom of at least one of the sulfonamide groups bound to hydrogen atoms or to an acyclic carbon atom to an acyclic carbon atom of a hydrocarbon radical substituted by carboxyl groups
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    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C311/00—Amides of sulfonic acids, i.e. compounds having singly-bound oxygen atoms of sulfo groups replaced by nitrogen atoms, not being part of nitro or nitroso groups
    • C07C311/30—Sulfonamides, the carbon skeleton of the acid part being further substituted by singly-bound nitrogen atoms, not being part of nitro or nitroso groups
    • C07C311/37—Sulfonamides, the carbon skeleton of the acid part being further substituted by singly-bound nitrogen atoms, not being part of nitro or nitroso groups having the sulfur atom of at least one of the sulfonamide groups bound to a carbon atom of a six-membered aromatic ring
    • C07C311/38—Sulfonamides, the carbon skeleton of the acid part being further substituted by singly-bound nitrogen atoms, not being part of nitro or nitroso groups having the sulfur atom of at least one of the sulfonamide groups bound to a carbon atom of a six-membered aromatic ring having sulfur atoms of sulfonamide groups and amino groups bound to carbon atoms of six-membered rings of the same carbon skeleton
    • C07C311/39—Sulfonamides, the carbon skeleton of the acid part being further substituted by singly-bound nitrogen atoms, not being part of nitro or nitroso groups having the sulfur atom of at least one of the sulfonamide groups bound to a carbon atom of a six-membered aromatic ring having sulfur atoms of sulfonamide groups and amino groups bound to carbon atoms of six-membered rings of the same carbon skeleton having the nitrogen atom of at least one of the sulfonamide groups bound to hydrogen atoms or to an acyclic carbon atom
    • C07C311/42—Sulfonamides, the carbon skeleton of the acid part being further substituted by singly-bound nitrogen atoms, not being part of nitro or nitroso groups having the sulfur atom of at least one of the sulfonamide groups bound to a carbon atom of a six-membered aromatic ring having sulfur atoms of sulfonamide groups and amino groups bound to carbon atoms of six-membered rings of the same carbon skeleton having the nitrogen atom of at least one of the sulfonamide groups bound to hydrogen atoms or to an acyclic carbon atom to an acyclic carbon atom of a hydrocarbon radical substituted by carboxyl groups
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    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C311/00—Amides of sulfonic acids, i.e. compounds having singly-bound oxygen atoms of sulfo groups replaced by nitrogen atoms, not being part of nitro or nitroso groups
    • C07C311/30—Sulfonamides, the carbon skeleton of the acid part being further substituted by singly-bound nitrogen atoms, not being part of nitro or nitroso groups
    • C07C311/45—Sulfonamides, the carbon skeleton of the acid part being further substituted by singly-bound nitrogen atoms, not being part of nitro or nitroso groups at least one of the singly-bound nitrogen atoms being part of any of the groups, X being a hetero atom, Y being any atom, e.g. N-acylaminosulfonamides
    • C07C311/46—Y being a hydrogen or a carbon atom
    • C—CHEMISTRY; METALLURGY
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    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C323/00—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups
    • C07C323/50—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups containing thio groups and carboxyl groups bound to the same carbon skeleton
    • C07C323/51—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups containing thio groups and carboxyl groups bound to the same carbon skeleton having the sulfur atoms of the thio groups bound to acyclic carbon atoms of the carbon skeleton
    • C07C323/60—Thiols, sulfides, hydropolysulfides or polysulfides substituted by halogen, oxygen or nitrogen atoms, or by sulfur atoms not being part of thio groups containing thio groups and carboxyl groups bound to the same carbon skeleton having the sulfur atoms of the thio groups bound to acyclic carbon atoms of the carbon skeleton with the carbon atom of at least one of the carboxyl groups bound to nitrogen atoms
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    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C327/00—Thiocarboxylic acids
    • C07C327/38—Amides of thiocarboxylic acids
    • C07C327/40—Amides of thiocarboxylic acids having carbon atoms of thiocarboxamide groups bound to hydrogen atoms or to acyclic carbon atoms
    • C07C327/44—Amides of thiocarboxylic acids having carbon atoms of thiocarboxamide groups bound to hydrogen atoms or to acyclic carbon atoms to carbon atoms of an unsaturated carbon skeleton
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    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C327/00—Thiocarboxylic acids
    • C07C327/38—Amides of thiocarboxylic acids
    • C07C327/48—Amides of thiocarboxylic acids having carbon atoms of thiocarboxamide groups bound to carbon atoms of six-membered aromatic rings
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    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D209/00—Heterocyclic compounds containing five-membered rings, condensed with other rings, with one nitrogen atom as the only ring hetero atom
    • C07D209/02—Heterocyclic compounds containing five-membered rings, condensed with other rings, with one nitrogen atom as the only ring hetero atom condensed with one carbocyclic ring
    • C07D209/04—Indoles; Hydrogenated indoles
    • C07D209/10—Indoles; Hydrogenated indoles with substituted hydrocarbon radicals attached to carbon atoms of the hetero ring
    • C07D209/14—Radicals substituted by nitrogen atoms, not forming part of a nitro radical
    • C07D209/16—Tryptamines
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D333/00—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom
    • C07D333/02—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom not condensed with other rings
    • C07D333/04—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom not condensed with other rings not substituted on the ring sulphur atom
    • C07D333/26—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom not condensed with other rings not substituted on the ring sulphur atom with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
    • C07D333/30—Hetero atoms other than halogen
    • C07D333/34—Sulfur atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D333/00—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom
    • C07D333/50—Heterocyclic compounds containing five-membered rings having one sulfur atom as the only ring hetero atom condensed with carbocyclic rings or ring systems
    • C07D333/52—Benzo[b]thiophenes; Hydrogenated benzo[b]thiophenes
    • C07D333/54—Benzo[b]thiophenes; Hydrogenated benzo[b]thiophenes with only hydrogen atoms, hydrocarbon or substituted hydrocarbon radicals, directly attached to carbon atoms of the hetero ring
    • C07D333/60—Radicals substituted by carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/02—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing at least one abnormal peptide link
    • C07K5/0215—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing at least one abnormal peptide link containing natural amino acids, forming a peptide bond via their side chain functional group, e.g. epsilon-Lys, gamma-Glu
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
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    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C2601/00—Systems containing only non-condensed rings
    • C07C2601/06—Systems containing only non-condensed rings with a five-membered ring
    • C07C2601/08—Systems containing only non-condensed rings with a five-membered ring the ring being saturated

Definitions

  • the present invention concerns amino acid derivatives possessing aspartyl protease inhibitory properties, in particular Ne-amino acid substituted L-lysine derivatives (and analogs) possessing aspartyl protease inhibitory properties. It describes also the synthetic methodology used to make these derivatives and their biological applications. In addition, this invention relates to different pharmaceutical compositions comprising these compounds.
  • the compounds and the pharmaceutical compositions of this invention have been shown to inhibit the activity of HIN aspartyl protease, an enzyme essential for virus maturation.
  • the inhibitory property may be advantageously used to provide compounds with antiviral properties against HIN viruses, including the HIN-1 and HIV-2 viruses.
  • HIN human immunodeficiency virus
  • AIDS immunodeficiency syndrome
  • HIN infection is characterized by a period immediately following infection, called asymptomatic, which is devoid- of clinical manifestations in the patient. Progressive HIN-induced destruction of the immune system then leads to increased susceptibility to opportunistic infections, which eventually produces a syndrome called AIDS-related complex (ARC) characterized by symptoms such as persistent generalized lymphadenopathy, fever, weight loss, followed itself by full blown AIDS.
  • ARC AIDS-related complex
  • the HIN-1 retrovirus attaches primarily to the CD4 receptor (a 58 kDa transmembrane protein) to gain entry into susceptible cells, through high- affinity interactions between the viral envelope glycoprotein (gp 120) and a specific region of the CD4 molecule found in CD4 (+) T-helper lymphocytes and other cells carrying the receptor (Lasky L.A. et al., Cell vol. 50, p. 975-985 (1987)).
  • the HIN genetic material in the form of R ⁇ A. is then transcribed into D ⁇ A by a viral enzyme carried by the virus called reverse transcriptase.
  • the viral D ⁇ A now called pro virus is then transported into the cell nucleus in the form of a preintegration complex and' attached to the cell D ⁇ A by another viral enzyme called integrase. Following integration, the viral D ⁇ A then serves as a template for viral gene expression by the host transcription system.
  • the primary R ⁇ A transcripts made from the provirus are synthesized by the host cell R ⁇ A.polymerase II whose activity is modulated by two virus-encoded proteins, Tat and Rev.
  • the viral proteins are expressed mainly in the form of polyproteins. After the infected cell has produced all the different HIV polyproteins and genetic material, they assemble at the cell membrane and are released from the cell in the form of an immature viral particle.
  • a third viral enzyme known as protease then cleaves the polyproteins to give the mature, infectious viral particle.
  • the polyproteins that are cleaved by the HIV protease are the Gag and Gag-Pol precursors, whose cleavage gives rise to structural proteins and viral enzymes.
  • a number of synthetic antiviral agents have been designed to block various stages in the replication cycle of HIV. Although only those developed against the viral enzymes have reached the market so far.
  • the latter include compounds which block viral reverse transcriptase (for example, didanosine and zidovudine (AZT)) 5 or the viral protease (for example, ritonavir and indinavir).
  • viruses have improved significantly the survival time and quality of life of AIDS patients, the administration of most of these agents leads to unwanted side effects, such as anemia, neurotoxicity and bone marrow suppression.
  • Anti-protease compounds represent the most-recent drugs developed to block HIV replication. These compounds inhibit the formation of infectious virions by interfering with the processing of viral polyprotein precursors by the viral protease.
  • the antiviral potential of HIV protease inhibitors was first demonstrated using peptidic inhibitors. Such peptidic compounds, however, are typically large and complex molecules that tend to exhibit poor bioavailability and limited stability in the body. New compounds devoid of these drawbacks are urgently needed to treat HIV infections.
  • mutations arising during HIV replication lead to resistance to the currently available protease inhibitors, so new compounds with original structure are desirable to fight these resistant viral strains.
  • the present invention provides a novel class of compounds, including their pharmaceutically acceptable derivatives. These compounds have an affinity for aspartyl proteases, in particular, HIV aspartyl protease. Therefore, these compounds are useful as inhibitors of such proteases. These compounds can be used alone or in combination with other therapeutic or prophylactic agents for the treatment or prophylaxis of viral infection.
  • the compounds of this invention are capable of inhibiting HIV viral replication in human CD4+ T-cells, by inhibiting the ability of HIV aspartyl protease to catalyse the hydrolysis of peptide bonds present in viral Gag and Gag-Pol polyproteins.
  • novel compounds can thus serve to reduce the production of infectious virions from acutely and chronically infected cells, and can inhibit the initial or further infection of host cells.
  • these compounds are useful as therapeutic and prophylactic agents to treat or prevent infection by HIV-1 and HIV-2, which may result in asymptomatic infection, AIDS- related complex (ARC), acquired immunodeficiency syndrome (AIDS), AIDS-related dementia, or similar diseases of the immune system, and related viruses such as HTLV-I and HTLV-II, and simian immunodeficiency virus.
  • HIV-1 and HIV-2 may result in asymptomatic infection, AIDS- related complex (ARC), acquired immunodeficiency syndrome (AIDS), AIDS-related dementia, or similar diseases of the immune system, and related viruses such as HTLV-I and HTLV-II, and simian immunodeficiency virus.
  • the present invention relates to a class of Ne-amino acid substituted L-lysine derivatives (including its lower and higher homologues and analogs) as well as their pharmaceutically acceptable derivatives (e.g. salts). Accordingly, the present invention in accordance with one aspect thereof provides a compound of formula I
  • n 3 or 4
  • Cx is selected from the group consisting of -COOM, COOR ⁇ , -CHO, -CH 2 OR 7 , - -CONHRg and -CONR, 0 R graffiti, wherein M is an alkali metal (e.g. Na, K, Cs, etc) or alkaline earth metal (Ca, Mg, etc.), wherein R, is selected from the group consisting of a straight or branched alkyl group of 1 to 6 carbon atoms, a cycloalkylalkyl group having 3 to 6 carbon atoms in the cycloalkyl part thereof and 1 to 3 carbon atoms in the alkyl part thereof,
  • M is an alkali metal (e.g. Na, K, Cs, etc) or alkaline earth metal (Ca, Mg, etc.)
  • R is selected from the group consisting of a straight or branched alkyl group of 1 to 6 carbon atoms, a cycloalkylalkyl group having 3 to 6 carbon
  • R- is selected from the group consisting of a benzenesulfonyl group of formula III,
  • R- is selected from the group consisting of H, a straight or branched alkyl group of 1 to 6 carbon atoms, a phenyl or a benzyl group
  • R. is selected from the group consisting of H, a group of formula Ilia
  • R 4C being selected from the group consisting of a straight or branched alkyl group of 1 to 6 carbon atoms, a cycloalkylalkyl group having 3 to 6 carbon atoms in the cycloalkyl part thereof and 1 to 3 carbon atoms in the alkyl part thereof, (e.g.
  • R 5 is selected from the group consisting of H, a straight or branched alkyl group of 1 to 8 carbon atoms, (e.g. CH 3 -, CH 3 CH 2 CH 2 - 5 CH 3 CH 2 CH 2 CH 2 - 5 iso-C hl 9 -, C 6 HvisorCH 2 -), HOCH,-, C 6 H 5 CH 2 OCH 2 -, benzyl-OCH(CH 3 ), HO 2 CCH 2 -, HO 2 CCH 2 CH 2> NC-CH 2 -, H 2 NC(O)CH,-, H,NC(0)CH 2 CH 2 -, 4-CH 3 C 6 H 4 CH 2 SCH 2 -, CH 3 SCH 2 CH 2 - , H 2 NCH 2 CH 2 CH 2 CH 2 -, C 6 H 5 -, C 6 H s CH 2 - ; C 6 H 5 CH(OH)-, C 6 H 5 CH(CN)-, C 6 F 5 CH 2 -, 4-(9-fluoreny
  • pyridine-2-CH 2 - C 5 H 4 N-3-CH 2 - (i.e. pyridine-3-CH 2 -), C 5 H 4 N- 4-CH 2 - (i.e. pyridine-4-CH 2 -).
  • Ra represents a member selected from the group consisting of
  • Met is a methylene linked to the ⁇ ' nitrogen
  • R 6 is selected from the group consisting of H, a straight or branched alkyl group of 1 to 4 carbon atoms and glycyl
  • R 7 is selected from the group consisting of H, a straight or branched alkyl group of 1 to 4 carbon atoms
  • R g is selected from the group consisting of a straight or branched alkyl group of 1 to 6 carbon atoms, a cycloalkyl group having 3 to 6 carbon atoms, a cycloalkylalkyl group having 3 to 6 carbon atoms in the cycloalkyl part and 1 to 3 carbon atoms in the alkyl part thereof,
  • R 9 is selected from the group consisting of H, a straight or branched alkyl group of 1 to 4 carbon atoms, -OH, -NH 2 and -CH 2 CH 2 OH
  • R 10 and R ⁇ are independently selected from the group consisting of H, a straight or branched alkyl group of 1 to 4 carbon atoms wherein m is 0 or 1
  • R, 2 and R )3 are independently selected from the group consisting of H, a straight or branched alkyl group of 1 to 4 carbon atoms, F, Cl, Br, I, -CF 3 , -NO 2 , -NR, 0 R ⁇ , -NHCOR 10 , ,- OR, 05 -OCH,C 6 H 5 , -SR ]0 , -COOR, 0 , -COR ]0 and -CH 2 OH, R, 0 and R n being as defined herein.
  • this invention provides a compound of formula IA
  • n 3 or 4
  • Cx is selected from the group consisting of -COOM, COOR ⁇ ,. -CHO, -CH 2 OR 7 , - CH 2 OCOR s , -CONHRp and -CONR 10 R, statement wherein M is an alkali metal (e.g. Na, K, Cs, etc) or alkaline earth metal,
  • R is selected from the group consisting of a straight or branched alkyl group of 1 to 6 carbon atoms, a cycloalkylalkyl group having 3 to 6 carbon atoms in the cycloalkyl part thereof and 1 to 3 carbon atoms in the alkyl part thereof, wherein R 2 is selected from the group consisting of a benzenesulfonyl group of formula III,
  • R 3 is selected from the group consisting of H, a straight or branched alkyl group of 1 to 6 carbon atoms, a phenyl or a benzyl group
  • R 4 is selected from the group consisting of H, C 6 H n - 5 C 5 H 10 N-CH 2 CH 2 - 5 OC 4 H 8 N- CH 2 CH 2 - (i.e. morpholine- 4-CH 2 CH 2 -) ; C 6 H 5 CH 2 CH 2 -, 2,3-(CH 3 O) 2 C 6 H 3 CH 2 -, C 6 H 5 -, 2-C 5 H 4 N, 3-C 5 H 4 N, 4-C 5 H 4 N, 3-quinolyl, CH 3 CO-, CF 3 CO, C 6 H 5 CO- 5 C 6 H 5 CS-, 4-CH 3 OC 6 H 4 CH 2 CO-, C 6 H 5 CH 2 CH 2 CO- ; iso-butyl-CO-, wo-propyl-CO-, te;-t-butyl-CO-, tert-butyl-CH 2 CO-, 1- pyrrolidine-CO-, 4-morpholine-CO-, carbotetrahydro-3-furanyloxy
  • R 5 is selected from the group consisting of H, CH 3 -, CH 3 CH 2 CH 2 -,CH 3 CH 2 CH 2 CH 2 -. wo-C 4 H 9 -, C 6 H n CH 2 -, HOCH 2 -, C 6 H 5 CH 2 OCH r benzyl-OCH(CH 3 ), HO 2 CCH 2 - 5 H0 2 CCH,CH,-, NC-CH 2 -, H 2 NC(O)CH 2 -, H 2 NC(O)CH 2 CH 2 -, 4-CH 3 C 6 H 4 CH 2 SCH 2 -, CH 3 SCH,CH 2 - H 2 NCH 2 CH 2 CH 2 CH 2 - 3 C 6 H 5 -, C 6 H 5 CH 2 -, C 6 F 5 CH 2 -, 4-tert-butyl-C 6 H 4 CH 2 - s 4- HOC 6 H 4 CH 2 -, 4-ber ⁇ zyl-O-C 6 H 4 CH 2 -, 4-NO 2 C 6 H 4 CH 2
  • F ⁇ is selected from the group consisting of H, a straight or branched alkyl group of 1 to 4 carbon atoms and glycyl
  • R 7 is selected from the group consisting of H, a straight or branched alkyl group of 1 to 4 carbon atoms
  • R 8 is selected from the group consisting of a straight or branched alkyl group of 1 to 6 carbon atoms, a cycloalkyl group having 3 to 6 carbon atoms, a cycloalkylalkyl group having 3 to 6 carbon atoms in the cycloalkyl part and 1 to 3 carbon atoms in the alkyl part thereof,
  • R- j is selected from the group consisting of H, a straight or branched alkyl group of 1 to 4 carbon atoms, -OH, - ⁇ H 2 and -CH 2 CH 2 OH
  • R, 0 and R are independently selected from the group consisting of H, a straight or branched alkyl group of 1 to 4 carbon atoms .
  • R 12 and R 13 are independently selected from the group consisting of H, a straight or branched alkyl group of 1 to 4 carbon atoms, F, Cl, Br, I, -CF 3 , -NO 2 , -NR 10 R ⁇ , -NHCOR 10 , - OR 10 , -OCH 2 C 6 H 5: -SR 10 , -COOR 10 , -COR 10 and -CH 2 OH, R 10 and R tile being as defined herein.
  • This invention also provides a compound of formula la
  • Cx is selected from the group consisting of -COOM and -CH 2 OH, M being an alkali metal or alkaline earth metal,
  • R 1A is selected from the group consisting of a straight or branched alkyl group of 1 to 6 carbon atoms, a cycloalkylalkyl group having 3 to 6 carbon atoms in the cycloalkyl part thereof and 1 to 3 carbon atoms in the alkyl part thereof,
  • R 2A and R 4A are independently selected from the group consisting of H, a straight or branched alkyl group of 1 to 4 carbon atoms, F, Cl, Br, I, -CF 3 , -NO 2 , -NR ]0 R,,, -NHCOR ]0 , - OR, 0 , -OCH,C 6 H 5J -SR 10 , -COOR 10 , -COR 10 and -CH 2 OH,
  • R is selected from the group consisting of H, a straight or branched alkyl group of 1 to 8 carbon atoms, (e.g. CH 3 -, CH 3 CH 2 CH 2 -,CH 3 CH 2 CH 2 CH 2 -, /.r ⁇ -C 4 H 9 -, C 6 H, ,CH 2 -) 5 HOCH,-, C 6 H 5 CH 2 OCH 2 -, benzyl -OCH(CH 3 ), HO 2 CCH 2 -, H0 2 CCH 2 CH 2 -, NC-CH 2 - 5 H 2 NC(O)CH 2 -, H 2 NC(0)CH 2 CH 2 -.
  • H a straight or branched alkyl group of 1 to 8 carbon atoms, (e.g. CH 3 -, CH 3 CH 2 CH 2 -,CH 3 CH 2 CH 2 CH 2 -, /.r ⁇ -C 4 H 9 -, C 6 H, ,CH 2 -) 5 HOCH,-, C 6 H 5
  • R 10 and R n are as defined herein.
  • This invention also provides a compound of formula lb
  • Cx is selected from the group consisting of -COOM, and -CH 2 OH, M being an alkali metal or alkaline earth metal,
  • R 1A is selected from the group consisting of a straight or branched alkyl group of 1 to 6 carbon atoms, a cycloalkylalkyl group having 3 to 6 carbon atoms in the cycloalkyl part thereof and 1 to 3 carbon atoms in the alkyl part thereof,
  • R 2A and R 4B are independently selected from the group consisting of H, a straight or branched alkyl group of 1 to 4 carbon atoms, F, Cl, Br, I, -CF 3 , -NO 2 , -NR 10 R n , -NHCOR, 0 , - OR )05 -OCH 2 C 6 H 5 , -SR 10 , -COOR 10 , -COR 10 and -CH 2 OH,
  • R 5A is selected from the group consisting of H, a straight or branched alkyl group of 1 to 8 carbon atoms, (e.g. CH 3 -, CH 3 CH 2 CH 2 -,CH 3 CH 2 CH 2 CH 2 - 5 iso-C U 9 -, C 6 H n CH 2 -), HOCH 2 -, C 6 H 5 CH 2 OCH , benzyl-OCH(CH 3 ), HO 2 CCH 2 -, HO 2 CCH 2 CH 2 -, NC-CH 2 -, H 2 NC(O)CH 2 -, H 2 NC(O)CH 2 CH 2 -, 4-CH 3 C 6 H 4 CH 2 SCH 2 -, H 2 NCH 2 CH 2 CH 2 CH 2 -, .
  • benzyl substituted by a group as defined for R 2A e.g.4-tert-butyl-C 6 H 4 CH 2 -, 4-HOC 6 H 4 CH 2 -, 4-benzyl- O-C 6 H 4 CH 2 -, 4-NO 2 C 6 H 4 CH 2 -, 2-FC 6 H 4 CH 2 -, 3-FC 6 H 4 CH 2 -, 4-FC 6 H 4 CH 2 -),
  • This invention also provides a compound of formula Ic
  • Cx is selected from the group consisting of -COOM and -CH 2 OH, M being an alkali metal or alkaline earth metal,
  • R IA is selected from the group consisting of a straight or branched alkyl group of 1 to 6 carbon atoms, a cycloalkylalkyl group having 3 to 6 carbon atoms in the cycloalkyl part thereof and 1 to 3 carbon atoms in the alkyl part thereof,
  • R 2B and R 4B are independently selected from the group consisting of H, a straight or branched alkyl group of 1 to 4 carbon atoms, F, Cl, Br, I, -CF 3 , -NO 2 , -NR 10 R, ) , -NHCOR )0 , - OR I0 , -OCH 2 C 6 H 5 , -SR, 0 , -COOR )0 , -COR 10 and -CH 2 OH,
  • R JA is selected from the group consisting of H, a straight or branched alkyl group of 1 to 8 carbon atoms, (e.g. CH 3 -, CH 3 CH 2 CH 2 -,CH 3 CH 2 CH 2 CH 2 -, zso-C 4 H 9 -, C 6 H U CH 2 -), HOCH 2 -, C 6 H 5 CH 2 OCH 2 -, benzyl-OCH(CH 3 ), HO 2 CCH 2 -, HO 2 CCH 2 CH,-, NC-CH,-, H 2 NC(O)CH 2 -.
  • H a straight or branched alkyl group of 1 to 8 carbon atoms
  • benzyl substituted by a group as defined for R 2A e.g.4-tert-butyl-C 6 H 4 CH 2 -, 4-HOC 6 H 4 CH 2 -, 4-benzyl- O-C 6 H 4 CH 2 -, 4- ⁇ O 2 C 6 H 4 CH 2 -, 2-FC 6 H 4 CH 2 -, 3-FC 6 H 4 CH 2 -, 4-FC 6 H 4 CH 2 -),
  • This invention also provides a compound of formula Id
  • Cx is selected from the group consisting of -COOM and -CH 2 OH, M being an alkali metal or alkaline earth metal,
  • R ]A is selected from the group consisting of a straight or branched alkyl group of 1 to 6 carbon atoms, a cycloalkylalkyl group having 3 to 6 carbon atoms in the cycloalkyl part thereof and 1 to 3 carbon atoms in the alkyl part thereof, wherein R 4A is selected from the group consisting of H.
  • R 4C is selected from the group consisting of a straight or branched alkyl group of 1 to 6 carbon atoms, a cycloalkylalkyl group having 3 to 6 carbon atoms in the cycloalkyl part thereof and 1 to 3 carbon atoms in the alkyl part thereof, (e.g.
  • R 5A is selected from the group consisting of H, a straight or branched alkyl group of 1 to 8 carbon atoms, (e.g. CH 3 -, CH 3 CH 2 CH 2 -,CH 3 CH 2 CH 2 CH 2 -, zso-C 4 H 9 -, C 6 H U CH 2 -), HOCH , C 6 H 5 CH 2 OCH 2 - 5 benzyl-OCH(CH 3 ), HO 2 CCH 2 -, HO 2 CCH 2 CH 2 -, NC-CH 2 -, H 2 NC(O)CH 2 -, H 2 NC(0)CH 2 CH 2 -, 4-CH 3 C 6 H 4 CH 2 SCH 2 -, CH 3 SCH 2 CH 2 - , H 2 NCH 2 CH 2 CH 2 CH 2 -, C 6 H 5 -, C 6 H 5 CH 2 -, C 6 H 5 CH(OH)-, C 6 H 5 CH(CN)-, C 6 F 5 CH 2 - 5 4-(9-fluor
  • pyridine-2-CH 2 - C 5 H 4 N-3-CH 2 - (i.e. pyridine-3-CH 2 -), C 5 H 4 N- 4-CH 2 - (i.e. pyridine-4-CH 2 -), 2-thiophene-CH 2 - > indole-3-CH 2 -, 2-benzothiophene-CH 2 -, N ⁇ - benzyi-imidazole-4-CH 2 - , imidazole-4-CH 2 - 5 thiazole-4-CH 2 - and substituted benzyl (e.g.
  • benzyl substituted by a group as defined for R 2A e.g.4-tert-butyl-C 6 H 4 CH 2 -, 4-HOC 6 H 4 CH 2 -, 4- benzyl-O-QH 4 CH 2 -, 4- ⁇ 0 2 C 6 H 4 CH,-, 2-FC 6 H 4 CH 2 -, 3-FC 6 H 4 CH 2 -, 4-FC 6 H 4 CH 2 -),
  • this invention provides pharmaceutical compositions in which these novel compounds of formula I or II, (as well as of formula IA, la, lb, Ic and Id) derived from L-lysine or L-lysine derivatives (as well as lower and higher homologues) are used to inhibit aspartyl proteases, including HIV aspartyl protease, thus providing protection against HIV infection.
  • the present invention provides a pharmaceutical composition
  • a pharmaceutical composition comprising a pharmaceutically acceptable carrier and a pharmaceutically effective amount of at least one compound of formula I, formula II (as well as of formulae IA, la, lb, Ic, and Id) and as applicable pharmaceutically acceptable ammonium salts thereof.
  • HIV protease and “HIV aspartyl protease” are used interchangeably and refer to the aspartyl protease encoded by the human immunodeficiency virus type 1 or 2. In a preferred embodiment of this invention, these terms refer to the human immunodeficiency virus type 1 aspartyl protease.
  • pharmaceutically effective amount refers to an amount effective in treating HIV infection in a patient.
  • prophylactically effective amount refers to an amount effective in preventing HIV infection in a patient.
  • patient refers to a mammal, including a human.
  • pharmaceutically acceptable carrier refers to a non-toxic carrier or adjuvant that may be administered to a patient, together with a compound of this invention, and which does not destroy the pharmacological activity 7 thereof.
  • the compounds of this invention include pharmaceutically acceptable derivatives of the compounds of formula I, formula II (as well as of formulae IA, la, lb, Ic, and Id) and as applicable pharmaceutically acceptable ammonium salts thereof.
  • a "pharmaceutically acceptable derivative” means any pharmaceutically acceptable salt, ester, or salt of such ester, of a compound of this invention or any other compound which, upon administration to a recipient, is capable of providing (directly or indirectly) a compound of this invention or an antivirally active metabolite or residue thereof.
  • Salts derived from appropriate bases include alkali metal (e.g., sodium), alkaline earth metal (e.g., magnesium), ammonium and N - (C M aIkyi) 4 + salts.
  • the compounds of this invention contain one or more asymmetric carbon atoms and thus may occur as racemates and racemic mixtures, single enantiomer, diastereomeric mixtures and individual diastereoisomers. All such isomeric forms of these compounds are expressly included in the present invention.
  • Each stereogenic carbon may be of the R or S configuration.
  • stable refers to compounds which possess stability sufficient to allow manufacture and administration to a mammal by methods known in the art. Typically, such compounds are stable at a temperature of 40°C or less, in the absence of moisture or other chemically reactive conditions, for at least a week.
  • Pharmaceutically acceptable salts of the compounds of this invention include those derived from pharmaceutically acceptable inorganic and organic acids and bases.
  • acid salts include: acetate, adipate, alginate, aspartate, benzoate, benzenesulfonate, bisulfate, butyratc, citrate, camphorate, camphorsulfonate, cyclopentanepropionate, digluconate, dodecylhydrogensulfate, dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptanoate, glycerophosphate, glycollate, hemisulfate, heptanoate, hexanoate, hydrochloride, hydrobromide, hydroiodide, 2-hydroxyethanesulfonate, lactate, maleate, malonate, methanesulfonate, 2- naphthylsulfonate, nicotinate, nit
  • This invention also envisions the quaternization of any basic nitrogen containing groups of the compounds disclosed herein.
  • the basic nitrogen can be quateraized with any agents known to those of ordinary skill in the art including, for example, lower alkyl halides, such as methyl, ethyl, propyl and butyl chlorides, bromides and iodides; dialkyl sulfates including dimethyl, diethyl, dibutyl and diamyl sulfates; long chain halides such as decyl, lauryl, myristyl and stearyl chlorides, bromides and iodides, and aralkyl halides including benzyl and phenethyl bromides. Water or oil-soluble or dispersible products may be obtained by such quaternization.
  • the compounds of this invention are readily prepared using conventional techniques from commercially available and inexpensive starting materials.
  • the relative ease of synthesis of the products described herein represents a marked advantage for the large scale preparation of these compounds.
  • the derivatives of the present invention may be readily obtained through sequences recognized by those knowledgeable in the art as straightforward. These sequences are presented in schemes 1 to 7 discussed below.
  • Scheme 1 illustrates a generic example for the preparation of a key intermediate needed for the synthesis of HIV protease inhibitors.
  • R represents an alkyl or cycloalkylalkyl side chain as defined above
  • R 2 represents a benzenesulfonyl group of formula III, a thiophenesulfonyl group of formula IV. a 1 -naphthylsulfonyl, a 2-naphthylsulfonyl or a 8-quinolinesulfonyl group as defined above
  • the N ⁇ ,N ⁇ -disubstituted L-lysine derivative 5 was obtained from commercially available L-lysine 1 in a four-step reaction sequence.
  • This preparation uses the cyclic form of L-lysine in order to manipulate the N -amino group without the need for protective groups.
  • L-lysine was transformed into L- -amino-e-caprolactam 2 upon treatment with hydrochloric acid in methanol followed by neutralization with sodium hydroxide.
  • the caprolactam 2 is also commercially available.
  • Reagents a) 1) MeOH/ H + (99.4%); 2) NaOMe, NH 4 CI, pH 11.5 (85%); b) Aldehyde, NaBH(OAc) 3 , DCE; c) arylsulfonyl chloride or substituted-arylsulfonyl chloride; TEA, CH 2 CI 2 , 3 h; d) 6N HCI, 12 h
  • Scheme 2 illustrates the preparation of HIV protease inhibitors bearing either a carboxylic function, compound 6, or an alcohol function, compound 8, on the final product.
  • this scheme shows the synthesis of a L-lysine derivative or a (25) 2,6-diaminohexanol derivative
  • R, and R 2 are as defined above b) R 3 represents H, a straight or branched alkyl group of 1 to 6 carbon atoms, a phenyl or a benzyl group c) R 4 is as defined above d) R 5 represents an amino acid side chain as defined above. Following the indications summarized in Scheme 2, derivative 5 is linked with a substituted amino acid using NN-carbonyldiimidazole as the activating reagent to yield derivative 6 in good to excellent yields.
  • N-acylated (or N-sulfonated) amino acids needed for the coupling reaction were prepared from the appropriate amino acid and acyl chloride (or sulfonyl chloride) using the Schotten-Baumen procedure.
  • derivative 5 is treated with trimethylsilyl chloride in methanol (HCl generated in situ) and the resulting methyl ester intermediate is reduced with lithium aluminum hydride (LAH) in THF to afford 7 in good yields.
  • LAH lithium aluminum hydride
  • the (2S) 2,6-diaminohexanol derivative 7 is linked to a substituted amino acid derivative as it is described above for the synthesis of derivative 6.
  • the L-lysine divative 6 can be further transformed into a variety of esters 9 as well as amide derivative 10 as shown in scheme 3. These transformations are done under standard reaction conditions. For example, the synthesis of ester 9 can be achieved upon activation of the acid 6 with DCC in the presence of a catalytic amount of NN-dimethylaminopyridine and an alcohol.
  • the amide 10 can be obtained as described earlier for the preparation of compound 6, see scheme 2.
  • R ]5 R 2 , R 3 , R 4 and R 5 are as described above
  • R 10 and R n same or different, represent an H or a straight or branched alkyl group of 1 to 4 carbon atoms.
  • Scheme 4 presents a second approach for the preparation of HIV protease inhibitors of formula 6 and 8. It proceeds by using commercially available Ne-benzyloxycarbonyl-L-lysine methyl ester hydrochloride (11) as the starting material. Reductive alkylation of derivative 11 with an appropriate aldehyde and sodium cyanoborohydride provided the derivative 12. Then, sulfonation with benzenesulfonyl chloride (or substituted-benzenesulfonyl chloride) in the presence of triethylamine (or diisopropylethylamine) in dichloromethane gave compound 13 in excellent yields for the two first steps.
  • Scheme 5 illustrates the preparation of an anti-protease derivative using a solid phase methodology in accordance with the present invention (see example 21). Any suitable solid phase substrate could be used in such preparation (K. Burgess, Solid phase organic synthesis, Wiley-Interscience, 2000).
  • R is w ⁇ -butyl
  • R 2 is 4-methylbenzenesulfonyl
  • R 3 , R 4 and R 5 are as described above
  • N ,N -disubstituted-L-lysine derivative such as 16
  • N -isobutyl-N -(4- methylbenzenesulfonyl)-Ne-(9-fluorenylmethoxycarbonyl)-L-lysine 16 is immobilized on &p- benzyloxybenzylalcohol resin (Wang resin) in DMF for a period of 16 h.
  • the resulting component 17 contained 0.28 meq. of L-lysine derivative / g of resin.
  • the resin can be coupled with a variety of N-acylated (or N-sulfonated) amino acids to give component 18.
  • the N-acylated (or N-sulfonated) amino acids are activated with N-hydroxysuccinimide and DCC in DMF. Cleavage of the resin with TFA in CH 2 C1 2 leads to the desired L-lysine derivative 19.
  • Any suitable solid phase support could be used, such as, for example, polystyrene (Ps see K. Burgess, Solid phase organic synthesis, Wiley-lnterscience, 2000.
  • LG Leaving group
  • Scheme 6 illustrates the preparation of substituted glycine derivatives used for the synthesis of several HIV protease inhibitors in accordance with the present invention (see examples 114 and 158 below for specific descriptions of the synthesis of such glycine derivatives):
  • N-phenylglycine 20 is treated with an excess butyllithium to give the dianion intermediate to which an appropriate alkyl halide (or arylalkyl halide or tosylate) is added and reacted for a period of 16 h.
  • the final products 21 are obtained in good to excellent yields.
  • An appropriate alkyl halide is defined as bearing a R 3 component which can sustain strong basic reaction conditions.
  • methyl bromoacetate 22 is treated with benzylamine in CH 2 C1 2 at room temperature for 16 h.
  • the N-benzylglycine methyl ester derivative 23 was obtained in 86% yield.
  • This intermediate can be either acylated with a carboxylic acid derivative and DCC in THF or sulfonatcd with an appropriate sulfonyl chloride and triethylamine in CH 2 C1 2 to give derivative 24 or 25 as desired in good to excellent yields.
  • LG Leaving group (Ci, Br.l, OTs)
  • R 4 Sulfonyl derivative Scheme 7 shows another methodology for the formation of Ne-substituted-glycyl-L-lysine HIV protease inhibitors via theNe-iodoacetyl-L-lysine derivative 26 (see example 105 for the detailed description of the synthesis of derivative 26 and its use).
  • N ⁇ ,N -disubstituted-L-lysine derivative 5 potassium sail is initially treated with chloroacctyl chloride in the presence of DIEA in THF to give the N ,N ⁇ -disubstituted-Ne-chloroacetyl-L-Iysine intermediate.
  • This intermediate is transformed into the iodoacetyl derivative 26 upon treatment with sodium iodide in dr>' acetone.
  • Compound 26 is then heated at reflux with a primary (or secondary) amine in the presence of of DIEA in THF to yield the desired Ne-substituted-glycyl-L-lysine derivative 6.
  • the compounds of this invention may be modified by appending appropriate functionalities to enhance selective biological properties.
  • modifications are known in the art and include those which increase biological penetration into a given biological system (e.g., blood, lymphatic system, central nervous system), increase oral availability, increase solubility to allow administration by injection, alter metabolism and alter rate of excretion.
  • the novel compounds of the present invention are excellent ligands for aspartyl proteases, particularly HIV-1 protease. Accordingly, these compounds are capable of targeting and inliibiting late stage events in the replication, i.e. the processing of the viral polyproteins by HIV encoded protease.
  • Compounds according to this invention advantageously inhibit the ability of the HIV-1 virus to infect immortalized human T cells over a period of days, as determined by an assay measuring the amount of extracellular p24 antigen ⁇ a specific marker of viral replication (see, Meek et al., Nature, 343, pp. 90-92 (1990)).
  • the compounds according to this invention may also be used as inhibitory or interruptive agents for other viruses which depend on aspartyl proteases, similar to HIV or HTLV aspartyl proteases, for obligatory events in their life cycle.
  • Such compounds inhibit the proteolytic processing of viral polyprotein precursors by inliibiting aspartyl protease. Because aspartyl protease is essential for the production of mature virions, inhibition of that processing effectively blocks the spread of virus by inhibiting the production and reproduction of infectious virions, particularly from acutely and chronically infected cells.
  • the compounds of this invention advantageously inhibit aspartyl proteases, thus blocking the ability of aspartyl proteases to catalyse the hydrolysis of peptide bonds.
  • the compounds of this invention may be employed in a conventional manner for the treatment or prevention of HIV, HTLV, and other viruses, which depend on aspartyl proteases for obligatory events in their life cycle. Such methods of treatment, their dosage levels and requirements may be selected by those of ordinary skill in the art from available methods and techniques.
  • a compound of this invention may be combined with a pharmaceutically acceptable adjuvant for administration to a virally infected patient in a pharmaceutically acceptable manner and in an amount effective to lessen the severity of the viral infection.
  • the compounds of this invention may be used in vaccines and methods for protecting individuals against viral infection over an extended period of time.
  • the compounds may be employed in such vaccines either alone or together with other compounds of this invention in a manner consistent with the conventional utilization of protease inhibitors in vaccines.
  • a compound of this invention may be combined with pharmaceutically acceptable adjuvants conventionally employed in vaccines and administered in prophylactically effective amounts to protect individuals over an extended period of time against viral infections, such as HIV infection.
  • the novel protease inhibitors of this invention can be administered as agents for treating or preventing viral infections, including HIV infection, in a mammal.
  • the compounds of this invention may be administered to a healthy or HlV-infected patient either as a single agent or in combination with other antiviral agents which interfere with the replication cycle of HIV.
  • the compounds of this invention may be administered with other antiviral agents which target different events in the viral life cycle, the therapeutic effect of these compounds is potentiated.
  • the co-administered antiviral agent can be one which targets early events in the viral life cycle, such as attachment to the cell receptor and cell entry, reverse transcription and viral DNA integration into cellular DNA.
  • Antiviral agents targeting such early life cycle events include among others polysulfated polysaccharides, sT4 (soluble CD4) and other compounds which block binding of virus to CD4 receptors on CD4 bearing T-lymphocytes and other CD4(+) cells, or inhibit fusion of the viral envelope with the cytoplasmic membrane, and didanosine (ddl), zalcitabine (ddC), stavudine (d4T), zidovudine (AZT) and lamivudine (3TC) which inhibit reverse transcription.
  • Other anti-retroviral and antiviral drugs may also be co- administered with the compounds of this invention to provide therapeutic treatment for substantially reducing or eliminating viral infectivity and the symptoms associated therewith.
  • antiviral agents examples include ganciclovir, dideoxycytidine, trisodium phosphonoformate, eflornithine, ribavirin, acyclovir, alpha interferon and trimenotrexate.
  • other types of drugs may be used to potentiate the effect of the compounds of this invention, such as viral uncoating inhibitors, inhibitors of Tat or Rev trans-activating proteins, antisense molecules or inhibitors of the viral integrase. These compounds may also be co- administered with other inhibitors of HIV aspartyl protease.
  • Combination therapies according to this invention exert a synergistic effect in inhibiting HIV replication because each component agent of the combination acts on a different site of HIV replication.
  • the use of such combinations also advantageously reduces the dosage of a given conventional anti-retro viral agent that would be required for a desired therapeutic or prophylactic effect as compared to when that agent is administered as a monotherapy .
  • These combinations may reduce or eliminate the side effects of conventional single anti-retroviral agent therapies while not interfering with the anti-retroviral activity of those agents.
  • These combinations reduce the potential of resistance to single agent therapies, while minimizing any associated toxicity.
  • These combinations may also increase the efficacy of the conventional agent without increasing the associated toxicity.
  • Preferred combination therapies include the administration of a compound of this invention with AZT, 3TC, ddl, ddC, d4T or other reverse transcriptase inhibitors.
  • the compounds of this invention may also be co-administered with other HIV protease inhibitors such as Ro 31-8959 (Saquinavir; Roche), L-735,524(Indinavir; Merck), AG- 1343 (Nelfinavir; Agouron), A-84538 (Ritonavir; Abbott), ABT-378/r (Lopinavir; Abbott), and VX-478 (Amprenavir; Glaxo) to increase the effect of therapy or prophylaxis against various viral mutants or members of other HIV quasi species.
  • HIV protease inhibitors such as Ro 31-8959 (Saquinavir; Roche), L-735,524(Indinavir; Merck), AG- 1343 (Nelfinavir; Agouron), A-84538 (Ritonavir; Abbott), ABT-378/r (Lopinavir; Abbott), and VX-478 (Amprenavir; Glaxo) to increase the effect of therapy or prophylaxis against various
  • the compounds of this invention can also be administered in combination with immunomodulators (e.g., bropirimine, anti-human alpha interferon antibody, IL-2, GM-CSF, methionine enkephalin, interferon alpha, diethyldithiocarbamate sodium, tumor necrosis factor, naltrexone and rEPO) antibiotics (e.g., pentamidine isethionate) or vaccines to prevent or combat infection and disease'associated with HIV infection, such as AIDS and ARC.
  • immunomodulators e.g., bropirimine, anti-human alpha interferon antibody, IL-2, GM-CSF, methionine enkephalin, interferon alpha, diethyldithiocarbamate sodium, tumor necrosis factor, naltrexone and rEPO
  • antibiotics e.g., pentamidine isethionate
  • vaccines to prevent or combat infection and disease'associated with HIV infection, such
  • compositions according to this invention may be comprised of a combination of an aspartyl protease inhibitor of this invention and another therapeutic or prophylactic agent.
  • the compounds of this invention can also be used as inhibitory agents for other viruses that depend on similar aspartyl proteases for obligatory events in their life cycle. These viruses include, but are not limited to, retroviruses causing AIDS-like diseases such as simian immunodeficiency viruses, HIV-2, HTLV-I and HTLV-II.
  • viruses include, but are not limited to, retroviruses causing AIDS-like diseases such as simian immunodeficiency viruses, HIV-2, HTLV-I and HTLV-II.
  • the compounds of this invention may also be used to inhibit other aspartyl proteases and, in particular, other human aspartyl proteases including renin and aspartyl proteases that process endothelin precursors.
  • compositions of this invention comprise any of the compounds of the present invention, and pharmaceutically acceptable salts thereof, with any pharmaceutically acceptable carrier, adjuvant or vehicle.
  • Pharmaceutically acceptable carriers, adjuvants and vehicles that may be used in the pharmaceutical compositions of this invention include, but are not limited to ion exchangers, alumina, aluminum stearate, lecithin, serum proteins, such as human serum albumin, buffer substances such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes, such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, cellulose-based substances, poiyethyleneglycol, sodium carboxymcthylccllulosc, polyacrylates, waxes, polycthylcne- polyoxyprop
  • compositions of this invention may be administered orally, parenterally by inhalation spray, topically, rectally, nasally, buccally, vaginally or via an implanted reservoir. We prefer oral administration or administration by injection.
  • the pharmaceutical compositions of this invention may contain any conventional non-toxic pharmaceutically acceptable carriers, adjuvants or vehicles.
  • parenteral as used herein includes subcutaneous, intracutaneous, intravenous, intramuscular, intra-articular, intrasynovial, intrasternal, intrathecal, intralesional and intracranial injection or infusion techniques.
  • the pharmaceutical compositions may be in the form of a sterile injectable preparation, for example, as a sterile injectable aqueous or oleaginous suspension.
  • This suspension may be formulated according to techniques known in the art using suitable dispersing or wetting agents (such as, for example, Tween 80) and suspending agents.
  • the sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent, for example, as a solution in 1,3-butanediol.
  • the acceptable vehicles and solvents that may be employed are amino acid, water, Ringer's solution and isotonic sodium chloride solution.
  • sterile, fixed oils are conventionally employed as a solvent or suspending medium.
  • any bland fixed oil may be employed including synthetic mono- or diglycerides.
  • Fatty acids, such as oleic acid and its glyceride derivatives are useful in the preparation of injectables, as are natural pharmaceutically-acceptable oils, such as olive oil or castor oil. especially in their polyoxyethylated versions.
  • These oil solutions or suspensions may also contain a long-chain alcohol diluent or dispersant, such as Ph. Helv. or a similar alcohol.
  • compositions of this invention may be orally administered in any orally acceptable dosage form including, but not limited to, capsules, tablets, and aqueous suspension and solutions.
  • carriers that are commonly used include lactose and com starch.
  • Lubricating agents such as magnesium stearate, are also typically added.
  • useful diluents include lactose and dried corn starch.
  • aqueous suspensions are administered orally, the active ingredient is combined with emulsifying and suspending agents. If desired, certain sweetening and/or flavoring and/or coloring agents may be added.
  • the pharmaceutical compositions of this invention may also be administered in the form of suppositories for rectal administration.
  • compositions can be prepared by mixing a compound of this invention with a suitable non-irritating excipient which is solid at room temperature but liquid at the rectal temperature and therefore will melt in the rectum to release the active components.
  • suitable non-irritating excipient include, but are not limited to, cocoa butter, beeswax, and polyethylene glycols.
  • Topical administration of the pharmaceutical compositions of this invention is especially useful when the desired treatment involves areas or organs readily accessible by topical application.
  • the pharmaceutical composition should be formulated with a suitable ointment containing the active components suspended or dissolved in a carrier.
  • Carriers for topical administration of the compounds of this invention include, but are not limited to, mineral oil, liquid petroleum, white petroleum, propylene glycol, polyoxyethylene or polyoxypropylene compound, emulsifying wax and water.
  • the pharmaceutical compositions can be formulated with a suitable lotion or cream containing the active compound suspended or dissolved in a carrier.
  • Suitable carriers include, but are not limited to, mineral oil, sorbitan monostearate, polysorbate 60, cety 1 esters wax cetearyl alcohol, 2-octyldodecanol, benzyl alcohol and water.
  • the pharmaceutical compositions of this invention may also be topically applied to the lower intestinal tracl by recta] suppository formulation or in a suitable neat formulation. Topically-transdermal patches are also included in this invention.
  • compositions of this invention may be administered by nasal aerosol or inhalation.
  • Such compositions are prepared according to teclmiques well-known in the art of pharmaceutical formulation and may be prepared as solutions in saline employing benzyl alcohol or other suitable preservatives, absorption promoters to enhance bioavailability, fluorocarbons, and/or other solubilizing or dispersing agents known in the art.
  • Dosage levels of between about 0.01 and about 25 mg/kg body weight per day, preferably between about 0.5 and about 25 mg/kg body weight per day of the active ingredient compound are useful in the prevention and treatment of viral infection, including HIV infection.
  • the pharmaceutical compositions of this invention will be administered from about 1 to about 5 times per day or alternatively, as a continuous infusion. Such administration can be used as a chronic or acute therapy.
  • the amount of active ingredient that may be combined with the carrier materials to produce a single dosage form will vary depending upon the patient treated and the particular mode of administration.
  • a typical preparation will contain from about 5%to about 95% active compound (w/w).
  • such preparations contain from about 20% to about 80% active compound.
  • a maintenance dose of a compound, composition or combination of this invention may be administered if necessary. Subsequently, the dosage or frequency of administration, or both, may be reduced, as a function of the symptoms, to a level at which the improved condition is retained. When the symptoms have been alleviated to the desired level, treatment should cease. Patients may, however, require intermittent treatment on a long-term basis, upon any recurrence of disease symptoms.
  • the compounds of this invention are also useful as commercial reagents which effectively bind to aspartyl proteases, particularly HIV aspartyl protease.
  • aspartyl proteases particularly HIV aspartyl protease.
  • the compounds of this invention, and their derivatives may be used to block proteolysis of a target peptide by an aspartyl protease, or may be derivatized to bind to a stable resin as a tethered substrate for affinity chromatography applications.
  • FRET fluorescence resonance energy transfer
  • Vi the enzyme velocity' in the presence of the inhibitory compound
  • [ I ] inhibitor concentration
  • [ S ] substrate concentration
  • Km - Michaelis-Menten constant and Ki app apparent Ki
  • Tables 1 and 2 were prepared by following Scheme 1, 2, 3, 4, 5, 6 or 7; the numbers of the compounds listed in the table correspond to the example numbers presented in the experimental section (see examples below). The activities of the compounds are also listed in the same tables demonstrating their potential usefulness.
  • Table 1 compounds of formula I wherem Y, n, Cx, R i5 R 2 , R 3 , R j and R 5 are as presented in Table 1.
  • Table 2 compounds of formula II wherem Y, n, Cx, R perpetrat R 2 , R 4 and ⁇ are as presented in Table 2.
  • HIV-1 Human immunodeficiency virus type 1
  • type 2 HOBt 1 -Hydroxybenzotriazole HPLC High performance liquid chromatography HTLV-I, -II Human T-cell lymphotropic virus type I, type II IL-2 lnterleukin-2
  • analytical plates can be treated with a 0.3% ninhydrin solution in ethanol containing 3% acetic acid and/or a CAM solution made of 20 g (NH 4 ) 6 Mo 7 O 24 and 8.3 g Ce(S0 4 ) 2 polyhydrate in water (750 mL) containing concentrated sulfuric acid (90 mL).
  • Preparative HPLC were perform on a Gilson apparatus equipped with a Cl 8 column, a215 liquid handler module and 15 mL/min capacity head pumps.
  • the HPLC is operated with a Gilson UniPoint System Software.
  • a solvent gradient was used starting from H 2 O/CH 3 CN (95%:5%) to 100% CH 3 CN over 25 min, and 100% CH 3 CN for a further 20 min to clean the column.
  • the organic phase was dried over MgSO 4 _ filtered and evaporated to an oil which crystallized on standing.
  • the solid was recrystallized from either dichloromethane, ether, hexanes or without solvent as indicated in each specific example.
  • the purity was evaluated by LC-MS and/or ⁇ NMR and was found to be ranging from 85 to 99%.
  • N ⁇ -isobutyl- ⁇ 7 ⁇ -(4-methylbenzenesulfonyl)-L-lysinehydrochloride (or other L-lysine derivative) (100 mg, 0.25 mmol) was weighed in the Bohdahn robotic reaction vessels. The solid was then added to 1 L 3.3M Cs 2 CO 3 solution to which 2 mL of THF was added. The tube was then stirred vigorously. The resulting mixture was treated with N-acylated (or N-sulfonated) amino acid (0.3 mmol) activated by NN-carbonyldiimidazole (0.3 mmol) dissolved in THF (1 mL). The stirring was continued for 2 h.
  • the resin was filtered and washed successively with DCM (100 mL, 2X), 1 :1 DCM : MeOH (100 mL, 3X), MeOH (50 mL, 2X) and ether (100 mL).
  • the resin was again swollen in DCM to which acetic anhydride (20 mL) was added. It was left to stand for 3 h and then filtered and washed as above.
  • the resulting resin was dried at room temperature in a dessicator in vac o.
  • the resulting resin (5.92 g) contained 0.28 mcq/g L-lysine derivative.
  • N-acylated (or N-sulfonated) amino acid was added to a solution of N-hydroxysuccinimide (0.5 mmol) and DCC (0.5 mmol) in DMF (3 mL).
  • the acid was activated for 3 h and filtered directly into the resin containing vessel.
  • the coupling reaction was allowed to proceed for 12 h at room temperature.
  • the resin was then washed successively with DCM, MeOH and ether as described above then dried in v ⁇ cuo.
  • N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-L -lysine potassium salt (or other L-lysine potassium salt derivative) (1 mmol) was weighed in a round bottom flask and was suspended in THF (20 L). This suspension was treated with a IN ⁇ aOH (1.5 mL) to pH 10. In another reaction vessel, a solution of N ⁇ protected amino acid (1 mmol) in THF (25 mL), was treated with 115 mg of N-h droxysuccinimide (1 mmol) and 206 mg DCC (1 mmol). The resulting mixture was stirred at room temperature for 4 h.
  • the L-lysine product (100 mg) was dissolved in MeC ⁇ (1 mL). The solution was added to 5 mL H 2 0 to form a turbid suspension. A IN ⁇ aOH solution (1 mol eq) was slowly added to ' the turbid suspension which became clear. The solution was frozen solid and lyophilized to yield a white powder (100%).
  • step C of this example The compound prepared in step C of this example (10.0 g, 51 mmol, free base) was dissolved in DCM (100 mL) and treated with diisopropylethylamine (10 mL) followed by freshly recrystallized 4-methylbenzenesulfonyl chloride (11.4 g, 57.3 mmol). The mixture was stirred overnight (TLC shows the reaction to be complete after 2 h). The solution was extracted with IN HCl and the organic layer was dried and evaporated. Then, the residue was dissolved in boiling CHC1 3 (5 mL), diluted with hexanes (200 mL) and placed in the refrigerator for 3 h. The precipitated product was filtered off and air dried giving 15.5 g of pure product.
  • Step E Preparation of N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-L-lysine hydrochloride ,
  • Step F Preparation of N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-Ne-[N' ⁇ -(4- methylbenzenesulfonyl)-L- ⁇ henylalanyl]-L-lysine
  • Step B Preparation of N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-Ne-[N' ⁇ -(4- methylbenzenesulfonyl)-L-ttyptophanyl]-L-lysine
  • the title compound was prepared from ⁇ ; c.-isobutyl-N -(4-methylbenzenesulfonyl)-L-lysine (150 mg, 0.42 mmol, example 1, step E) as described in general procedure Be using N -(4- methylbenzenesulfonyl)-L-tryptophan (180 mg, 0.5 mmol) which was prepared in step A of this example.
  • the final product was triturated with ether to yield 180 mg (69%) of the desired material.
  • Step B Preparation of N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-Ne-[N' ⁇ -(4- acetamidobenzenesulfonyl)-L-phenylalanyl]-L-lysine
  • the title compound was prepared from N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-L-lysine (100 mg, 0.29 mmol. example 1, step E) as described in general procedure Be using N -(4- acetamidobenzenesulfonyl)-L-phenylalanine (102 mg, 0.29 mmol) which was prepared in step A of this example.
  • the final product was triturated with ether to yield 101 mg (57%) of the desired material.
  • Step B Preparation of N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-Ne-(N ⁇ -benzenesulfonyl-L- tryptophanyl)-L-lysine
  • the title compound was prepared from N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-L-ly sine (1.0 g, 2.9 mmol, example 1 , step E) as described in general procedure Be using N ⁇ -benzenesulfonyl- L-tryptophan (1.72 g, 5 mmol) which was prepared in step A of this example.
  • the final product was triturated with ether to yield 1 J g of the crude material. Purification of 500 mg of the crude material by HPLC gave 322 mg (64%) of pure adduct.
  • Step B Preparation of N ⁇ -isobuty'l-N ⁇ -(4-nitrobenzenesulfonyl)-L- ⁇ -amino-e-caprolactam N ⁇ -isobutyl-L- ⁇ -amino-e-caprolactam (example 1, step C) (4.14 g, 21.1 mmol, free base) was dissolved in DCM (50 mL) and treated with diisopropylethylamine (6.0 mL, 30 mmol) followed by freshly recrystallized 4-nitrobenzenesulfonyl chloride (5.09 g, 21 J mmol). The mixture was stirred overnight (TLC shows the reaction to be complete after 2 h).
  • Step D Preparation of N ⁇ -isobutyl-N ⁇ -(4-aminobenzenesulfonyl)-Ne-[N' ⁇ -(4- aminobenzenesulfonyl)-L-tryptophanyl]-L-lysine
  • the title compound was prepared from N ⁇ -isobutyl-N ⁇ -(4-nitrohenzenesulfonyl)-L-lysine hydrochloride (200 mg, 0.52 mmol, step C) as described in general procedure Be using N ⁇ -(4- nitrobenzenesulfonyl)-L-tr ⁇ ptophan (300 mg, 0.8 mmol) which was prepared in step A of this example.
  • the intermediate derivative was reduced following the indications of general procedure E.
  • the final product was purified by HPLC to give 101 mg (53%) of pure adduct. !
  • the title compound was prepared from ⁇ r ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-L-lysine (100 mg, 0.29 mmol, example 1, step E) as described in general procedure Be using N ⁇ -(4- nitrobenzenesulfonyl)-L-tryptophan (120 mg, 0.3 mmol) which was prepared in step A of example 5.
  • the final product was purified by HPLC to give 66 mg (36%) of pure adduct.
  • Step B Preparation of N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-Ne-[N' ⁇ -(4- methylbenzenesulfonyl)-D-phenylalanyl]-L-lysine
  • the title compound was prepared from N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-L-lysine (100 mg, 0.29 mmol, example 1 , step E) as described in general procedure Be using N -(4- methylbenzenesulfonyl)-D-phenylalanine (160 mg, 0.5 mmol) which was prepared in step A of this example.
  • the final product was triturated with ether to yield 49 mg (29%) of the desired material.
  • Step B Preparation of N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-Ne-(N' ⁇ -benzenesulfonyl-L- phenylalanyl)-L-lysine
  • the title compound was prepared from N ⁇ -isobutyl- ⁇ f ⁇ -(4-methylbenzenesulfonyl)-L-Iysine hydrochloride (120 mg, 0.34 mmol, example 1, step E) as described in general procedure Be using N ⁇ -benzenesulfonyl-L-phenylalanine (300 mg, 1.0 mmol) prepared in step A of this example.
  • the final product was purified by preparative HPLC to yield 107 mg (56%) of the desired material.
  • Step B Preparation of N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-Ne-[N' ⁇ -(4- chlorobenzenesulfonyl)-L-phenylalanyl]-L-lysine
  • the title compound was prepared from N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-L-lysine (100 mg, 0.29 mmol, example 1, step E) as described in general procedure Be using Na-(4- chlorobenzenesulfonyl)-L-phenylalanine (89 mg, 0.3 mmol) prepared in step A of this example.
  • the final product was purified by preparative HPLC to yield 56 mg (33%) of the desired material.
  • Step A Preparation of N ⁇ -(4-nitrobenzenesulfonyl)-L-phenylalanine L-phenylalanine was reacted with 4-nitrobenzenesulfonyl chloride under the conditions used in general procedure A giving the title compound which was recrystallised in ether (37%).
  • Step B Preparation of N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-Ne-[N' ⁇ -(4- nitrobenzenesulfonyl)-L-phenylalanyl]-L-lysine
  • the title compound was prepared fromN ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-L-lysine (100 mg, 0.29 mmol, example 1, step E) as described in general procedure Be using N ⁇ -(4- nitrobenzenesulfonyl)-L-phenylalanine (125 mg, 0.5 mmol) prepared in step A of this example.
  • the final product was purified by preparative HPLC to yield 91 mg (52%) of the desired material.
  • Step B Preparation of N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-Ne-[N' ⁇ -(4- methylbenzenesulfonyl)-L-tyrosyl]-L-lysine
  • the title compound was prepared from N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-L-lysine (100 mg, 0.29 mmol, example 1 , step E) as described in general procedure Be using N ⁇ -(4- methylbenzenesulfonyl)-L-tyrosine (160 mg, 0.5 mmol) prepared in step A of this example.
  • the final product was purified by preparative HPLC to yield 78 mg (46%) of the desired material.
  • the title compound was prepared from N ⁇ -isobutyl-N ⁇ -(4-methylber ⁇ zenesulfonyl)-L-lysine hydrochloride (200 mg, 0.59 mmol, example 1 , step E) as described in general procedure Be using N ⁇ -(4-nitrobenzenesulfonyl)-L-tryptophan (300 mg, 0.75 mmo ⁇ ) prepared in step A of example 5.
  • the intermediate, N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-Ne-[N' -(4- nitrobenzenesulfonyl)-L-tryptophanyl]-L-]ysine was reduced following the conditions of general procedure E.
  • the final product was purified by preparative HPLC to yield 266 mg (58%) of the • desired material.
  • Step B Preparation of N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-Ne-[N' -(4- nitrobenzenesulfonyl)-L-alanyl]-L-lysine
  • the title compound was prepared from N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-L-lysine hydrochloride (100 mg, 0.29 mmol, example 1, step E) as described in general procedure Be using N ⁇ -(4-nitrobenzenesulfonyl)-L-alanine (140 mg, 0.5 mmol) prepared in step A of this example.
  • the final product was purified by preparative HPLC to yield 10 mg (6.5%) of the desired material.
  • Step B Preparation of N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-Ne-[N ' ⁇ -(benzenesulfonyl)- L-norvalyl]-L-lysine
  • the title compound was prepared from N ⁇ -isobutyl-Ntx-(4-methylbenzenesulfonyl)-L-ly sine (100 mg, 0.29 mmol, example 1, step E) as described in general procedure Be using N ⁇ - benzenesulfonyl-L-norvalinc (120 mg, 0.5 mmol) prepared in step A of this example.
  • the final product was purified by preparative HPLC to yield 15 mg (10%) of the desired material.
  • Step B Preparation ofN ⁇ -isobutyl- ⁇ 7 ⁇ -(4-methylbenzenesulfonyl)-Ne-[N' -(benzenesulfonyl)- L-norleucyl]-L-lysine
  • the title compound was prepared from N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-L-lysine (100 mg. 0.29 mmol, example 1, step E) as described in general procedure Be using N ⁇ - benzenesulfonyl-L-norleucine (125 mg, 0.5 mmol) prepared in step A of this example.
  • the final product was purified by preparative HPLC to yield 13 mg (8.5%) of the desired material.
  • Step B Preparation of N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-Ne-[N' ⁇ -(4- nitrobenzenesulfonyl)-L-leucyl]-L-lysine
  • the title compound was prepared fromN ⁇ -isobutyl-No!-(4-methylbenzenesulfonyl)-L-lysine (100 mg, 0.29 mmol, example 1, step E) as described in general procedure Be using N ⁇ -(4- nitrobenzenesulfonyl)-L-leucine (150 mg, 0.5 mmol) prepared in step A of this example.
  • the final product was purified by preparative HPLC to yield 28 mg (17%) of the desired material.
  • Step B Preparation of ⁇ -isobutyl-N ⁇ -(4-methyIbenzenesulfonyl)-Ne-[N' ⁇ -(benzenesulfonyl)- 4-tr-7/M-hydroxy-L- ⁇ rolyl]-L-lysine
  • the title compound was prepared fromN ⁇ -isobutyl-7 ⁇ -(4-methylbenzenesulfonyl)-L-]ysine (100 mg, 0.29 mmol, example 1, step E) as described in general procedure Be using N ⁇ - benzenesulfonyl-4-tr ⁇ n.s'-hydroxy-L-proline (130 mg, 0.5 mmol) prepared in step A of this example.
  • the final product was purified by preparative HPLC to yield 9 mg (6%) of the desired material.
  • Step A Preparation of N ⁇ -(4-fluorobenzenesulfonyl)-L-phenylalanine L-phenylalanine was reacted with 4-fluorobenzenesulfonyl chloride under the conditions used in general procedure A giving the title compound which was recrystallised in ether (40%).
  • Step B Preparation of N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-Ne-[N' -(4- fluorobenzenesulfonyl)-L-phenylalanyl]-L-lysine
  • the title compound was prepared fromN ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-L-lysine (100 mg, 0.29 mmol, example 1, step E) as described in general procedure Be using N -(4- fluorobenzenesulfonyl)-L-phenylalanine (160 mg, 0.5 mmol) prepared in step A of this example.
  • the final product was purified by preparative HPLC to yield 87 mg (52%) of the desired material.
  • Step B Preparation ofN ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-Ne-[N' ⁇ -(2-naphthylsulfony3)- L-phenylalanyl]-L-lysine
  • the title compound was prepared from N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-L-ly sine (100 mg, 0.29 mmol, example 1, step E) as described in general procedure Be using N ⁇ -(2- naphfhylsulfonyl)-L-phenylalanine (175 mg, 0.5 mmol) prepared in step A of this example.
  • the final product was purified by preparative HPLC to yield 55 mg (32%) of the desired material.
  • Step B Preparation of N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-Ne-[N' ⁇ -(4- bromobenzenesulfonyl)-L-phenylalanyl]-L-lysine
  • the title compound was prepared from N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-L-ly sine (100 mg, 0.29 mmol, example 1, step E) as described in general procedure Be using N ⁇ -(4- bromobenzenesulfonyl)-L-phenylalanine (190 mg, 0.5 mmol) prepared in step A of this example.
  • the final product was purified by preparative HPLC to yield 56 mg (30%) of the desired material.
  • Step B Preparation of N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-Ne-(9- fluorenylmethoxycarbonyl)-L-lysine
  • the title compound was prepared from N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-L-lysine by following the indications of general example Be using commercially available N-(9- fluorenylmethoxycarbonyloxy) succinimide instead of the usual combination of reactants i.e. N- protected amino acid, N-hydroxysuccinimide and DCC.
  • Step C Preparation of N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-Ne-[N' ⁇ -(4- methylbenzenesulfonyl)-glycyl]-L-lysine
  • step B The title compound was prepared from solid phase bound N ⁇ -isobutyl-N ⁇ -(4- methylbenzenesulfonyl)-Ne-(9-fluorenylmethoxycarbonyl)-L-lysine (step B) as described in general procedure Bb using N ⁇ -(4-methylbenzenesulfonyl)-glycine (110 mg, 0.5 mmol) prepared in step A of this example.
  • the final product was purified by preparative HPLC to yield 30 mg (42%) of the desired material.
  • Step B Preparation of Na-isobutyl-Na-(4-methylbenzenesulfonyl)-Ne-(A r 'a-benzenesulfonyl-L- leucyl]-L-lysine
  • the title compound was prepared from solid phase bound 7 ⁇ -isobutyl-N ⁇ -(4- methylbenzenesulfonyl)-Ne-(9-fluorenylmethoxycarbonyl)-L-lysine as described in general procedure Bb using N ⁇ -benzenesulfonyl-L-leucine (130 mg, 0.5 mmol) prepared in step A of this example.
  • the final product was purified by preparative HPLC to yield 30 mg (39%) of the desired material.
  • Step B Preparation of N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-Ne-[N' ⁇ -(4- trifluoromethylbenzenesulfonyl)-L-phenylalanyl]-L-lysine
  • the title compound was prepared from solid phase bound N ⁇ -isobutyl-N ⁇ -(4- methylbenzenesulfonyl)-Ne-(9-fluorenyImethoxycarbonyl)-L-lysine as described in general procedure Bb using N ⁇ -(4-trifluoromethylbenzenesulfonyl)-L-phenylalanine (180 mg, 0.5 mmol) prepared in step A of this example. The final product was purified by preparative HPLC to yield 50 mg (56%) of the desired material.
  • the title compound was prepared from N ⁇ -isobuty'l-N ⁇ -(4-methylbenzenesulfonyl)-L-lysine hydrochloride (250 mg, 0.61 mmol, example 1, step E) as described in general procedure Be using N ⁇ -(2-thiophenesulfonyl)-L-phenylalar ⁇ ine (155 mg, 0.5 mmol) which was prepared in step A of this example.
  • the crude material was purified by HPLC to give 272 mg (66%) of pure adduct.
  • Step B Preparation ofN-t-isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-Ne-(N' ⁇ -benzenesulfonyl)-L- asparagyl]-L-lysine
  • the title compound was prepared from solid phase bound N ⁇ -isobutyl-N ⁇ -(4- methylbenzenesulfonyl)-Ne-(9-fluorenylmethoxycarbonyl)-L-lysine as described in general procedure Bb using N ⁇ -benzenesulfonyl-L-asparagine (350 mg, 1.0 mmol) prepared in step A of this example.
  • the final product was purified by preparative HPLC to yield 70 mg (91%) of the desired material.
  • Step B Preparation of N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-Ne-[N' ⁇ -(4- methylbenzenesuIfonyI)-L-4-nitrophenylaIanyl]-L-Iysine
  • the title compound was prepared from solid phase bound N ⁇ -isobutyI-N ⁇ -(4- methylbenzenesulfonyl)-Ne-(9-fluorenylmethoxycarbonyl)-L-lysine as described in general procedure Bb using N ⁇ -(4-methylbenzenesulfonyl)-L-4-nitrophenylalanine (180 mg, 0.5 mmol) prepared in step A of this example.
  • the final product was purified by preparative HPLC to yield 25 mg (28%) of the desired material.
  • Step B Preparation of N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-Ne-[N' ⁇ -(4- methylbenzenesulfonyl)-L-phenylglycyl]-L-lysine
  • the title compound was prepared from solid phase bound N ⁇ -isobutyl-N ⁇ -(4- methylbenzenesulfonyl)-Ne-(9-fluorenylmethoxycarbonyl)-L-lysine as described in general procedure Bb using N ⁇ -(4-methylbenzenesulfonyl)-L-phenylglycine (150 mg, 0.5 mmol) prepared in step A of this example.
  • the final product was purified by preparative HPLC to yield 55 mg (68%) of the desired material.
  • Step A Preparation of N ⁇ -isobutyl-N ⁇ -(4-nitrobenzenesulfonyl)-Ne-(9- fluorenylmethoxycarbonyl)-L-lysine
  • Step B Preparation of N ⁇ -isobutyl-N ⁇ -(4-nitrobenzenesulfonyl)-Ne-[N' ⁇ -(4- acetamidobenzenesulfonyl)-L-phenylalanyl]-L-lysine
  • the title compound was prepared from solid phase bound N ⁇ -isobutyl-N ⁇ -(4- nitrobenzenesulfonyl)-Ne-(9-fluorenylmethoxycarbonyl)-L-lysine (step A) as described in general procedure Bb using N ⁇ -(4-acetamidobenzenesulfonyl)-L-phenylalanine (400 mg, 1.2 mmol) which was prepared in step A of example 3.
  • the final product was purified by preparative HPLC to yield 55 mg (60%) of the desired material.
  • the title compound was prepared from solid phase bound N ⁇ -isobutyl-N ⁇ -(4- nitrobenzenesulfonyl)-Ne-(9-fluorenylmethoxycarbonyl)-L-lysine as described in general procedure Bb using N ⁇ -(2-thiophenesulfonyl)-L-phenylalanine (300 mg, 1.2 mmol) which was prepared in step A of example 24.
  • the final product was purified by preparative HPLC to yield 46 mg (54%o) of the desired material.
  • Step A Preparation of N ⁇ -acetyl-L-phenylalanine L-phenylalanine was reacted with acetyl chloride under the conditions used in general procedure A giving the title compound which was recrystallised from ether (97%>). This compound is also commercially available.
  • Step B Preparation of N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-Ne-(N' ⁇ -acetyl-L- phenylalanyl)-L-lysine
  • the title compound was prepared from N ⁇ -isob ⁇ tyd-N ⁇ -(4-methylbenzenesulfonyl)-L-lysine (150 mg, 0.42 mmol, example 1, step E) as described in general procedure Be using N ⁇ -acetyl-L- phe ylalanine (207 mg, 1.0 mmol) which was prepared in step A of this example.
  • the final product was purified by preparative HPLC ' to yield 121 mg (59%) of the desired material.
  • the title compound was prepared from solid phase bound N ⁇ -isobutyl-N ⁇ -(4- methylbenzenesulfonyl)-Ne-(9-fluorenylmethoxycarbonyl)-L-lysine as described in general procedure Bb using commercially available N ⁇ -benzyloxy carbonyl -L-phenylalanine (300 mg, 1.2 mmol).
  • the final product was purified by preparative HPLC to yield 33 mg (37%) of the desired material.
  • Step B Preparation of N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-Ne-[N' ⁇ -(4- methyIbenzenesulfonyl)-L-seryl]-L-lysine
  • the title compound was prepared from solid phase bound N ⁇ -isobutyl-N ⁇ -(4- methy]benzenesulfonyl)-Ne-(9-fluorenylmethoxycarbonyl)-L-lysine as described in general procedure Bb using N ⁇ -(4-mclhyIbcnzcncsulfonyl)-L-scrine (150 mg, 1.2 mmol) prepared in step A of this example. The final product was purified by preparative HPLC to yield 35 mg (46%) of the desired material.
  • the title compound was prepared from solid phase bound N ⁇ -isobutyl-N ⁇ -(4- methylbenzenesulfonyl)-Ne-(9-fluorenylmethoxycarbonyl)-L-lysine as described in general procedure Bb using N ⁇ -(4-methyIbenzenesulfonyl)-L-cyclohexylalanine (350 mg, 1.2 mmOl) prepared in step A of this example.
  • the final product was purified by preparative HPLC to yield 19 mg (22%o) of the desired material.
  • Step B Preparation of • N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-Ne-[N' ⁇ -(4- methylbenzenesulfonyl)-L-glutaminyl]-L-lysine
  • the title compound was prepared from solid phase bound N ⁇ -isobutyl-N ⁇ -(4- methylbenzenesulfonyl)-Ne-(9-fluorenylmethoxycarbonyl)-L-lysine as described in general procedure Bb using N ⁇ -(4-methylbenzenesulfonyl)-L-glutamine (360 mg, 1.2 mmol) prepared in step A of this example.
  • the final product was purified, by preparative HPLC to yield 18 mg (21%) of the desired material.
  • Step B Preparation of N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-Ne-[N' ⁇ -(4- methylbenzenesulfonyl)-L-2-thiophenylalanyl]-L-lysine
  • the title compound was prepared from solid phase bound N ⁇ -isobutyl-N ⁇ -(4- methylbenzenesulfonyl)-Ne-(9-fluorenylmethoxycarbonyl)-L-lysine as described in general procedure Bb using N ⁇ -(4-methylbe:nzenesulfonyl)-L-2-thiophenylalanine (390 mg, 1.2 mmol) prepared in step A of this example. The final product was purified by preparative HPLC to yield 15 mg (18%) of the desired material.
  • the title compound was prepared from solid phase bound N ⁇ -isobutyl-N ⁇ -(4- methylbenzenesulfonyl)-Ne-(9-fluorenylmethoxycarbonyl)-L-lysine as described in general procedure Bb using commercially available N ⁇ -(9-fluorenylmethoxycarbonyl)-L-serine (390 mg, 1.2 mmol).
  • the final product was purified by preparative HPLC to yield 15 mg (18%) of the desired material.
  • the title compound was prepared from solid phase bound N ⁇ -isobutyl-N ⁇ -(4- methylbenzenesulfonyl)-Ne-(9-fluorenylmethoxycarbonyl)-L-lysine as described in general procedure Bb using commercially available N ⁇ -(9-fluorenylmethoxycarbonyl)rL- cyclohexylalanine (470 mg, 1.2 mmol). The final product was purified by preparative HPLC to yield 11 mg (12%) of the desired material.
  • Step B Preparation of N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-Ne-[N' ⁇ -(4- methylbenzenesulfonyl)-L-glutamyl]-L-lysine
  • the title compound was prepared from solid phase bound N ⁇ -isobutyl-N ⁇ -(4- methylbenzenesulfonyl)- ⁇ 7 e-(9-fluorenylmethoxycarbonyl)-L-lysine as described in general procedure Bb using N ⁇ -(4-methylbenzenesulfonyl)-L-glutamic acid (360 mg, 1.2 mmol) prepared in step A of this example.
  • the final product was purified by preparative HPLC to yield 20 mg (25%) of the desired material.
  • Ne-terr-butoxycarbonyl-L-lysine was reacted with 4-methylbenzenesulfonyl chloride under the conditions used in general procedure A giving the title derivative which was used without purification in the next step.
  • Step B Preparation of N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-Ne-[N' ⁇ -(4- methylbenzenesulfonyl)-L-lysyl]-L-lysine
  • the title compound was prepared from solid phase bound N ⁇ -isobutyl-N ⁇ -(4- methylbenzenesulfonyl)-Ne-(9-fluorenylmethoxycarbonyl)-L-lysine as described in general procedure Bb using N ⁇ -(4-methylbenzenesulfonyl)-Ne-te7"/-butoxycarbonyl-L-lysine (360 mg, 1.2 mmol) prepared in step A of this example.
  • the final product was purified by preparative HPLC to yield 10 mg (12%) of the desired material.
  • the title compound was prepared from solid phase bound N ⁇ -isobutyl-N ⁇ -(4- methylbenzenesulfonyl)-Ne-(9-fluorenylmethoxycarbonyl)-L-lysine (200 mg, 0.1 mmol) in a similar fashion to general procedure Bb using commercially available N ⁇ -(9- fluorenylmethoxycarbonyl)-O-benzyl-L-serine (100 mg, 0.24 mmol) with DCC (100 mg, 0.48 mmol) and HOBt (50 mg, 0.37 mmol) as the activating reagents.
  • the intermediate adduct was again deprotected and coupled with 4-methylbenzenesulfonyl chloride in CH 2 C1 2 before being cleaved from the resin with TFA.
  • the final product was purified by preparative HPLC to yield 30 mg (42%)) of the desired material.
  • Step B Preparation of N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-Ne-[N' ⁇ -(4- methylbenzenesulfonyl)-L-aspartyl]-L-lysme
  • the title compound was prepared from solid phase bound N ⁇ -isobutyl-N ⁇ -(4- methylbenzenesulfonyl)- ⁇ 7 e-(9-fluorenylmethoxycarbonyl)-L-lysine as described in general procedure Bb using N ⁇ -(4-methylbenzenesulfonyl)-L-aspartic acid (340 mg, 1.2 mmol) prepared in step A of this example.
  • the final product was purified by preparative HPLC to yield 20 mg (26%>) of the desired material.
  • Step B Preparation of N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-Ne-[N' ⁇ -(4- methylbenzenesulfonyl)-L-3-(2-thianaphthyl)-alanyl]-L-lysine
  • the title compound was prepared from solid phase bound N ⁇ -isobutyl-N ⁇ -(4- methylbenzenesulfonyl)-Ne-(9-fluorenylmethoxycarbonyl)-L-lysine as described in general procedure Bb using 4-methylbenzenesulfonyl-L-3-(2-thianaphthyl)-alanine (450 mg, 1.2 mmol) prepared in step A of this example.
  • the final product was purified by preparative HPLC to ield 25 mg (28%) of the desired material.
  • the title compound was prepared from N ⁇ -isobutyl-N ⁇ -(4-nitrobenzenesulfonyl)-L-lysine hydrochloride (350 mg, 1.0 mmol, example 5, step C) as described in general procedure Be using N ⁇ -(4-methylbenzenesulfonyl)-L-tryptophan (335 mg, 1.1 mmol) which was prepared in step A of example 2.
  • the title compound was prepared from N ⁇ -isobutyl-N ⁇ -(4-nitrobenzenesulfonyl)-L-lysine (385 mg, 1.0 mmol, example 5, step C) as described in general procedure Be using N ⁇ - benzenesulfonyl-L-phenylalanine (335 mg, 1.1 mmol) which was prepared in step A of example 8.
  • the final product was purified by preparative HPLC to yield 550 mg (85%) of N ⁇ -(4- nitrobenzenesulfonyl)-N ⁇ -isobutyl-Ne-(N' ⁇ -benzenesulfonyl)-L-phenylalanyl)-L-lysine.
  • the latter derivative (200 mg) was hydrogenolysed following the indications of general procedure E. Purification by HPLC gave the desired material (129 mg, 65%).
  • the title compound was prepared from N ⁇ -isobutyl-N ⁇ -(4-nitrobenzenesulfonyl)-L-lysine hydrochloride (400 mg, 1.03 mmol, example 5, step C) as described in general procedure Be using N ⁇ -(2-thiophenesulfonyl)-L-phenylalanine (311 mg, 1.2 mmol) which was prepared in step A of example 24.
  • Step B • Preparation of N ⁇ -(4-aminobenzenesulfonyI)-N ⁇ -isobutyl-Ne-[N' ⁇ -(4- methylbenzenesulfonyl)-L-asparagyl]-L-lysine
  • the title compound was prepared from N ⁇ -isobutyl-N ⁇ -(4-nitrobenzenesulfonyl)-L-lysine hydrochloride (400 mg, 1.03 mmol, example 5, step C) as described in general procedure Be using N ⁇ -(4-methylbenzenesulfonyl)-L-asparagine (286 mg, 1.0 mmol) which was prepared in step A this example.
  • Example 48 Preparation of ⁇ r ⁇ -isobutyI-N ⁇ -(4-methyIbenzenesuIfonyl)-Ne-(N' ⁇ - benzyloxycarbonyI-L-asparagyI)-L-lysine
  • the title compound was prepared from solid phase bound N ⁇ -isobutyl-N ⁇ -(4- methylbenzenesulfonyl)- ⁇ ? e-(9-fluorenylmethoxycarbonyl)-L-lysine as described in general procedure Bb using commercially available N ⁇ -benzyloxycarbonyl-L-asparagine (319 mg, 1.2 mmol).
  • the final product was purified by preparative HPLC to yield 46 mg (41%) of the desired material.
  • Step B Preparation of N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)- ⁇ / e-(N' ⁇ -benzoyl-L- phenylalanyl)-L-lysine
  • the title compound was prepared from solid phase bound N ⁇ -isobutyl-N ⁇ -(4- methylbenzenesulfonyl)-Ne-(9-fluorenylmethoxycarbonyl)-L-lysine as described in general procedure Bb using N ⁇ -benzoyl-L-phenylalanine (321 mg, 1.2 mmol). The final product was purified by preparative HPLC to yield 44 mg (58%) of the desired material.
  • Step B Preparation of N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-Ne-[N' ⁇ -(4- morpholinecarbonyl)-L-phenylalanyl]-L-lysine
  • the title compound was prepared from solid phase bound N ⁇ -isobutyl- ⁇ r ⁇ -(4- methylbenzenesulfonyl)-Ne-(9-fluorenylmethoxycarbonyl)-L-lysine as described in general procedure Bb usingN ⁇ -(4-morpholinecarbonyl)-L-phenylalanine (330 mg, 1.2 mmol).
  • the final product was purified by preparative HPLC to yield 41 mg (53%) of the desired material.
  • Step A Preparation of N ⁇ -pivaloyl-L-phenylalanine L-phenylalanine was reacted with pivaloyl chloride under the conditions used in general procedure A giving the title compound which was recrystallised from hexanes (40%>).
  • the title compound was prepared from solid phase bound N ⁇ -isobutyl-N ⁇ -(4- methylbenzenesulfonyl)-Ne-(9-fluorenylmethoxycarbonyl)-L-lysine as described in general procedure Bb using N ⁇ -pivaloyl-L-phenylalanine (300 mg, 1.2 mmol).
  • the final product was purified by preparative HPLC to yield 49 mg (66%) of the desired material.
  • Step B Preparation of N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-Ne-[N' ⁇ -(2- thiophenesulfonyl)-L-tryptophanyI]-L-lysine
  • step E as described in general procedure Be using N ⁇ -(2- thiophenesulfonyl)-L-tryptophan (90 mg, 0.3 mmol) which was prepared in step A of this example.
  • the final product was triturated with ether to yield 12 mg (6%>) of the desired material.
  • Example 54 Preparation of N ⁇ -isobutyI-N ⁇ -(2-thiophenesuIfonyI)-Ne-[N , ⁇ -(4- methylbenzenesuIfonyI)-L-phenylalanyl]-L-Iysine
  • N ⁇ -isobutyl-L- ⁇ -amino-e-caprolactam (example 1, step C) (2.56 g, 14.0 mmol, free base) was dissolved in DCM (50 mL) and treated with diisopropylethylamine (4.0 mL, 20 mmol) followed by freshly recrystaUized 2-thiophenenesulfonyl chloride (2.56 g, 14.0 mmol). The mixture was stirred overnight (TLC shows the reaction to be complete after 2 h). The solution was extracted with IN HCl and the organic layer was dried and evaporated. Then, the residue was dissolved in boiling MeOH (150 mL) and placed in the refrigerator for 3 h. The thin needles obtained were filtered off and air dried giving 3.6 g (78%) of pure product. This compound was used without further purification in the next step.
  • Step C Preparation of N ⁇ -isobutyl-N ⁇ -(2-thiophenesulfonyl)-Ne-[N ' ⁇ -(4- methylbenzenesulfonyl)-L-phenylalanyl]-L-lysine
  • N ⁇ -isobutyl-N ⁇ -(2-thiophenesulfonyl)-L-lysine hydrochloride 150 mg, 0.5 mmol
  • THF 20 mL
  • a IN ⁇ aOH 0.5 mL
  • the title compound was prepared in a three-step sequence from rert-butyl bromoacetate. Initially, tert-butyl bromoacetate (1.91 g, 10 mmol) dissolved in isobutylamine (60 mmol) was stirred at room temperature for 2 h. The reaction mixture was filtered and the excess isobutylamine was distilled yielding 80%> of pure N-isobutyl gl cine tert-butyl ester.
  • step D the intermediate was reacted with 4-methylbenzenesulfonyl chloride (8 mmol) as described for the preparation of Na- isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-L- ⁇ -amino-e-caprolactam presented in example 1 (step D).
  • 4-methylbenzenesulfonyl chloride 8 mmol
  • triethylamine was used instead of diisopropylethylamine .
  • deprotection of the tert-butyl ester with TFA provided the final product quantitatively (two last steps).
  • the title compound was prepared from N ⁇ -isobutyl-N ⁇ -(4-methy lbenzenesulfonyl)-L-lysine (100 mg, 0.29 mmol, example 1 , step E) as described in general procedure Be using N ⁇ -isobutyl-N ⁇ - (4-methylbenzenesulfonyl)-glycine (90 mg, 0.3 mmol) which was prepared in step A of this example. The final product was triturated with ether to yield 40 mg (21%) of the desired material.
  • the title compound was prepared from N ⁇ -(4-aminobenzenesulfonyl)-N ⁇ -isobutyI-L-Iysine (100 mg, 0.29 mmol, example 166, step B) as described in general procedure Be using N ⁇ -(4- acetamidobenzenesulfonyl)-L-phenylalanine (110 mg, 0.3 mmol) which was prepared in step A of example 3.
  • the final product was purified by HPLC to give 50 mg (23%) of pure adduct.
  • the title compound was prepared by treating N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-Ne- [N' ⁇ -(2-thiophenesulfonyl)-L-phenylalanyl]-L-lysine (60 mg, 0.1 mmol, example 24) dissolved in MeOH (2 mL) with DCC (1 eq.). The reaction mixture was stirred at room temperature for a period of 2 h. Filtration and evaporation of the solvent followed by HPLC purification gave the desired methyl ester (15 mg, 22%).
  • the title compound was prepared fromN ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-L-lysine (100 mg, 0.29 mmol, example 1, step E) as described in general procedure Be using N ⁇ -(4- methylbenzenesulfonyl)-L-asparagine (90 mg, 0.3 mmol) which was prepared in step A of example 47.
  • the final product was triturated with ether to yield 95 mg (15%) of the desired material.
  • Step B Preparation of N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-Ne-(N' ⁇ -pivaloyl-L- asparagyl)-L-lysine
  • the title compound was prepared from N ⁇ -isobutyI-N ⁇ -(4-methyIbenzenesulfonyl)-L-ly sine (100 mg, 0.29 mmol, example 1 , step E) as described in general procedure Be using N ⁇ -pivaloyl-L- asparagine (65 mg, 0.3 mmol) which was prepared in step A this example.
  • the final product was purified by preparative HPLC to yield 21 mg (12%) of the desired material.
  • Step A Preparation of N ⁇ -benzoyl-L-asparagine ' .
  • Step B Preparation of ⁇ 7 ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-Ne-(N' ⁇ -benzoyl-L- ' asparagyl)-L-Iysine ,
  • the title compound was prepared from N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-L-lysine (100 mg, 0.29 mmol, example 1, step E) as described in general procedure Be using N ⁇ -benzoyl-L- asparagine (70 mg, 0.3 mmol) which was prepared in step A this example.
  • the final product was purified by preparative HPLC to yield 14 mg (9%>) of the desired material.
  • Step B Preparation of N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-Ne-benzyloxycarbonyl-L- lysine methyl ester ; ' ⁇ i . ⁇ .
  • Step G Preparation of N ⁇ -isobutyl-N ⁇ -(4-methy lbenzenesulfonyl)-Ne-[l, 2,3,4- i ⁇ • ' ⁇ • tetrahydroisoquinoline-N , -(4-methylbenzenesulfonyl)-3-carbonyl]-L-lysine x ⁇ • .
  • the final product was prepared from N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-Ne-[l, 2,3,4- tetrahydroisoquinoline-3 -carbonyl]-L- lysine trifluoroacetic acid salt (step F above) following the indications of step B of this example.
  • the desired material was obtained in 71%) yield.
  • the title compound was prepared fromN ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-Ne-[N' ⁇ -(4- ' ⁇ ! i ⁇ methylbenzenesulfony l)-L-phenylalanyl]-L-lysine methyl ester, product of example 68, following the indications of general procedure F.
  • the thioamide was Obtained in 64%) yield.
  • the title compound was prepared from N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-Ne-(N' ⁇ - benzenesulfonyl-L-tryptophanyl)-L-lysine, product of example 4, following the indications of general procedure F.
  • the thioamide was obtained in 34% yield.
  • the title compound was prepared from N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-Ne-(N' ⁇ - benzoyl-L-phenylalanyi)-L-ly sine methyl ester, product of example 70, following the indications of general procedure F.
  • the thioamide was obtained in 81 %> yield.
  • the title compound was prepared from N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-L-lysine hydrochloride (1.2 g, 3.1 mmol, example 1, ' step E) suspended in THF (50 mL) and l ⁇ ⁇ aOH (0.5 mL. to reach pH 10) as described in general procedure Be using commercially available N ⁇ - . benzyloxycarbonyl-L-phenylalanine (1.0 g, 3.5 mmol), N-hydroxysuccinimide (0.4 g, 3.5 mmol) and DCC (1.1 g, 4.8 mmol). The final product was triturated with ether to yield 1.61 g (95%) of the desired material.
  • This product was prepared from N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-Ne-(N' ⁇ -tert- butoxycarbonyl-L-phenylalanyl)-L-lysine (600 mg, 1.0 mmol, example 78) as described in general procedure C. The residue was triturated with ether and placed under high vacuum to yield a hard white foam (440 mg, 88%). ' '
  • Example 80 Preparation of N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-Ne-[N , ⁇ - (carbotetrahydro-3-furanyloxy)-L-phenylalanyl]-L-lysine
  • the title compound was prepared fromN ⁇ -isobufyl-N ⁇ -(4-methylbenzenesulfonyl)-L-lysine (100 mg, 0.29 mmol, example 1, step E) as described in general procedure Be using N ⁇ - (carbotetrahydro-3-fliranyloxy)-L-phenylalanine (84 mg, 0.3 mmol) which was prepared in step A of this example.
  • the final product was triturated with ether to yield 51 mg (27%>) of the desired material.
  • the title compound was prepared fromN ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-L-lysine (100 mg, 0.29 mmol, example 1, step E) as described in general procedure Be using commercially available N ⁇ -tert-butoxycarbonyl-N ⁇ -methyl-L-phenylalanine (83 mg, 0.3 mmol).
  • the final product was triturated with ether to yield 176 mg (90%>) of the desired material.
  • Example 82 Preparation of N ⁇ -isoburyl-N ⁇ -(4-methylbenzenesulfonyl)-Ne-(N , ⁇ -/'er - butoxycar,bonyl-L-methionyl)-L-lysine
  • the title compound was prepared from N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-L-lysine (100 mg, 0.29 mmol, example 1, step E) as described in general procedure Be using commercially available N ⁇ -tert-butoxycarbonyl-L-methionine (75 mg, 0.3 mmol). The final product was triturated with ether to yield 170 mg (96%) of the desired material.
  • the title compound was prepared fromN ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-L-lysine (100 mg, 0.29 mmol, example 1, step E) as described in general procedure Be using commercially available N ⁇ -tert-butoxycarbonyl-S-(4-methylbenzyl)-L-cysteine (67 mg, 0.3 mmol). The final product was triturated with ether to yield 130 mg (65%) of the desired material.
  • the title compound was prepared from N ⁇ -isobutyl-N ⁇ -(4-methylbenze ! nesulfonyl)-L-lysine (100 mg, 0.29 mmol, example 1, step E) as described in general procedure Be using commercially available N ⁇ -tert-butoxycarbonyl-O-benzyl-L-threonine (93 mg, 0.3 mmol). The final product was triturated with ether to yield 195 mg (98%) of the desired material.
  • the title compound was prepared from N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-L-lysine (100 mg, 0.29 mmol, example 1, step E) as described in general procedure Be using commercially available ⁇ -tert-butoxycarbonyl-N ⁇ -benzyl-L-liistidine (100 mg, 0.3 mmol). The final product was triturated with ether to yield 133 mg (65%) of the desired material.
  • the title compound was prepared fromN ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-L-lysine (100 mg, 0.29 mmol, example 1, step E) as described in general procedure Be using commercially available N ⁇ -tert-butoxycarbonyl-L-tryptophan (90 mg, 0.3 mmol). The final product was triturated with ether to yield 157 mg (81%) of the desired material.
  • the title compound was prepared fromN ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-L-lysine (100 mg, 0.29 mmol, example 1, step E) as described in general procedure Be using commercially available N ⁇ -tert-butoxycarbonyl-O-benzyl-L-tyrosine (111 mg, 0.3 mmol). The final product was triturated with ether to yield 145 mg (68%) of the desired material.
  • This product ' was prepared from N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-Ne-(N' ⁇ -tert- butoxyca ⁇ -bonyl-N J ⁇ -methyl-L-phenylalanyl)-L-lysine (100 mg, 0.16 mmol, example 81) as described in general procedure C. The residue was triturated with ether and placed under high vacuum to yield a hard white foam (25 mg, .30%).
  • This product was prepared from N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-Ne-(N' ⁇ -tert-- butoxycarbonyl-L-methionyl)-L-lysine (95 rrig, 0.16 mmol, example 82) as described in general procedure C. The residue was triturated with ether and placed under high vacuum to- yield a hard white foam (40 mg, . 51 %) .
  • This product was prepared from N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-Ne-[N' ⁇ -tert- butoxycarbonyl-S-(4-methylbenzyl)-L-cysteinyl]-L-lysine (91 mg, 0.16 mmol, example 83) as described in general procedure C. The residue was triturated with ether and placed under high vacuum to yield a hard white foam (60 mg, 66%>).
  • This product was prepared from N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-Ne-(N' ⁇ -tert- butoxycarbonyl-O-benzyl-L-threonyl)-L-lysine (110 mg, 0.18 mmol, example 84) as described in general procedure C. The residue was triturated with ether and placed under high vacuum to yield a hard white foam (40 mg, 41%).
  • This product was prepared from N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-Ne-(N' ⁇ -tert- butoxycarbonyl-N ⁇ -benzyl-L-histidinyl)-L-lysine (110 mg, 0.16 mmol, example 85) as described in general procedure C. The residue was triturated with ether and placed under high vacuum to yield a hard white foam (51 mg, 54%).
  • This product was prepared from N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-Ne-(N' ⁇ -tert- butoxycarbonyl-L-tryptophanyl)-L-lysine (100 mg, 0.15 mmol, example 86) as described in i general procedure C. The residue was triturated with ether and placed under high vacuum to yield a hard white foam (27 mg, 33%):
  • This product was prepared from N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-Ne-(N' ⁇ -tert- butoxycarbonyl-O-benzyl-L-tyrosyl)-L-lysine (115 mg, 0.16 mmol, example 87) as described in general procedure C. The residue was triturated with ether and placed under high vacuum to yield a hard white foam ' (19 mg, 19%).
  • Step B Preparation , ⁇ of N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-Ne-(N' ⁇ -acetyl-L- tryptophanyl)-L-lysine [ ⁇ ⁇ '
  • the title compound was prepared from N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-L-lysine hydrochloride (400 mg, 1.0 mmol, example 1 , step E) suspended in THF (20 mL) and IN ⁇ aOH (0.5 mL, to reach pH 10) as described in general procedure Be using N ⁇ -acetyl-L-tryptophan (300 mg, i .4 mmol) prepared in step A of this example, N-hydroxysuccinimide (115 mg, 1.0 mmol) and DCC (210 mg, 1.0 mmol). The crude material (600 mg) was purified by preparative HPLC to yield 411 mg (45%>) of the desired material.
  • Step B Preparation of N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-Ne-(N' ⁇ -pivaloyl-L- tryptophanyl)-L-lysine •
  • the title ' compound was prepared from N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-L-lysine hydrochloride (400 mg, 1.0 mmol, example 1, step E) suspended in THF (20 mL) and I ⁇ aOH (0.5 mL, to reach pH 10) as described in general procedure Be using N ⁇ -pivaloyl-L-tryptophan (350 mg, 1.5 mmol) prepared in step A of this example, N-hydroxysuccinimide (115 mg, 1.0 mmol) and DCC (210 mg, 1.0 mmol). The crude material (488 mg) was purified by preparative HPLC to yield 244 mg (50%) of the desired material.
  • N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-Ne-(N' ⁇ -tert-butoxycarbonyl-L-phenylalanyl)-L- lysine (50 mg, 0.08 mmol, example 78) is added to a mixture of of trifluoroacetic anhydride and TFA and was stirred for a period of 30 min. Afterwards, the reactant were evaporated and the residue triturated with ether to give 40 mg (74%) of the title compound.
  • Example 104 Preparation of N ⁇ -isobutyI-N ⁇ -(4-methyIbenzenesuIfonyI)-Ne-(N' ⁇ - benzenesulfonyl-L-tryptophanyl)-L-lysine 2,3-dihydroxypropyl ester
  • the title compound was prepared from N ⁇ -isobutyl-N ⁇ -(4-methyIbenzenesulfonyl)-L-lysme potassium salt (2.0 g, 5.1 mmol) suspended in a mixture of THF (20 mL), DIEA (1.5 mL, 8.6 mmol) and chloroacetyl chloride (0.8 mL, 10.1. mmol). The reaction mixture was stirred for a period of 30 min. Then, a solution of 2 ⁇ HCl (20 mL) was added and the solution extracted with EtOAc (25 mL, 3X). The organic phase was dried with MgSO 4 , filtered and evaporated to a brown oil. The crude material was purified by flash chromatography using a solvent gradient from 39: 1 to 19: 1 CH 2 Cl 2 /MeOH. The product was isolated as . a yellowish oil (1 J g, 78% yield).
  • Step B Preparation: of N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-Ne-iodoacetyl-L-lysine
  • the title ' compound was prepared from N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-Ne- iodoacetyl-L-lysine: (200 mg, 0.38 mmol, example 105, step B) by following the indications of general procedure H using DIEA (0.19 mL, 1.09 mmol) and 3-aminopyridine (175 mg, 1.36 mmol). The crude material was purified by preparative HPLC. The product was isolated as a
  • the title compound was prepared from N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-Ne- iodoacetyl-L-l sine (150 mg, 0.29 mmol, example 105, step B) by following the indications of general procedure H using DIEA (0.14 mL, 0.81 mmol) and 2,3-dimethoxybenzylamine (0.12 mL, 0.81 mmol). The crude material was purified by preparative HPLC. The product was isolated as a solid (25 mg, 16% yield). • '
  • the title compound was prepared from N ⁇ -isobutyl-N ⁇ -(4-methyIbenzenesulfonyl)-Ne- iodoacetyl-L-lysine (200 mg, 038 mmol, example 105, step B) by following the indications of general procedure H using DIEA (0.19 mL, 1.09 mmol) and 2-aminopyridine (170 mg, 1.31 mmol). The crude material was purified by preparative HPLC. The product was isolated as a solid (88 mg, 47% yield).
  • the title compound was prepared from N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-Ne- iodoacetyi-L-lysine (200 mg, 0.38 mmol, example 105, step B) by following the indications of general procedure H using DIEA (0.20 mL, 1.10 mmol) and benzylamine (320 mg, 2.99 mmol).
  • DIEA 0.20 mL, 1.10 mmol
  • benzylamine 320 mg, 2.99 mmol
  • the crude material was purified by preparative HPLC.
  • The. product was isolated as a solid (137 mg, 71% yield). ;
  • the title compound was prepared from N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-Ne- iodoacetyl-L-lysine (150 mg, 0.29 mmol, example 105, step B) by following the indications of general procedure H using DIEA (0.12 mL, 0.69 mmol) and l-(2-aminoethyl)piperidine (0.12 mL, 0.84 mmol). The crude material was purified by preparative HPLC. The product was isolated as a solid (65 mg, 33% yield).
  • the title compound was prepared from N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-Ne- iodoacetyl-L-lysine (150 mg, 0.29 mmol, example 105, step B) by following the indications of general procedure H using DIEA (0.12 mL, 0.69 mmol) and 4-(2-aminoethyl)morpholine (0.11 mL, 0.84' mmol). The crude material was purified by preparative HPLC. The product was isolated as a solid (51 mg, 34% yield). ;
  • the title compound was prepared from N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-Ne- iodoacetyl-L-lysine (150 mg, 0.29 mmol, example 105, step B) by following the indications of general procedure H using DIEA (0.10 mL, 0.57 mmol) and 4-aminopyridine (76 mg, 0.59 mmol).
  • DIEA 0.10 mL, 0.57 mmol
  • 4-aminopyridine 76 mg, 0.59 mmol
  • the title compound was prepared from N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-Ne- iodoacetyl-L-Iysine (200 mg, 0.38 mmol, example 105, step B) by following the indications of general procedure H using DIEA (0.19 mL, 1.09 mmol) and 3-aminoquinoline (260 mg, 1.80 mmoi). The crude material was purified by preparative HPLC. The product was isolated as a solid (24 mg, 12% yield).
  • N-phenylglycine (1.51 g, 10.0 mmol) dissolved. in THF (20 mL) was treated with butyllithium 2.5M in hexane (8.8 mL, 22.0 mmol) under an inert atmophere of argon at ' - 78 °C. The mixture was stirred for 15 min before the addition of benzylbromide (1.3 mL, 1 1 mmol). The reaction mixture was stirred for 2 h while warming up to room temperature.. Then, 2 ⁇ HCl ' (25 mL) was added and the mixture extracted with EtOAc (30 mL, 3X).
  • the title compound was prepared from solid phase bound N ⁇ -isobutyl-N ⁇ -(4- methylbenzenesulfonyl)-Ne-(9-fluorenylmethoxycarbonyl)-L-lysine (200 mg) in asimilar fashion to general procedure Bb using N-benzyl-N-phenylglycine (100 mg, 0.45 mmol) prepared in step A of this example with DCC (200 mg, 0.97 mmol) and HOBt (100 mg, 0.74 mmol) as the activating reagents.' The final product was purified by preparative HPLC to yield 21 mg (75%>) of the desired material. ;
  • the title compound was prepared from N-phenylglycine (200 mg, 1.3 mmol) as described for the preparation of N-benzyl-N-phenylglycine (example 114, step A) using methyl iodide (1 mL, 6.6 mmol) instead of benzylbromide.
  • the crude material was purified by preparative HPLC. The product was isolated as a solid (90 mg, 41% yield).
  • Step B • Preparation of N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-Ne-(N' ⁇ -methyl- ⁇ ⁇ - phenylglycyl)-L-lysine ' .
  • the title compound was prepared from solid phase bound N ⁇ -isobutyl-N ⁇ -(4- methylbenzenesulfonyl)-Ne-(9-fluorenylmethoxycarbonyl)-L-lysine (200 mg) ' in a similar fashion to general procedure Bb using N-methyl-N-phenylglycine (75 mg, 0.45.mmol) prepared in step A of this example with DCC (200 mg, 0.97 mmol) and HOBt (100 mg, 0.74 mmol) as the activating reagents.
  • the final product was purified by preparative HPLC to yield 7 mg (25%o) of the desired material.
  • Step B Preparation of (2S,2'S) 2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N-[N' ⁇ -(4- methylbenzenesulfonyl)-S-tryptophanyl]-2,6-diaminohexanol
  • the title product was prepared from (2S) .2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6- diaminohexanol (400 mg, 1.2 mmol, step A) following the indications of general procedure Bd using N ⁇ -(4-methylbenzenesulfonyl)-L-tryptophan (450 mg, 1.3 mmol, example 2, step A), EDC (290 mg, 1.5 mmol) and HOBt (100 mg, 1.5 mmol) .
  • the crude material was purified by flash chromatography using a solvent gradient from 39:1 to 19:1 CH 2 Cl 2 /MeOH. The product was isolated as a yellowish oil (420 mg, 48% yield).
  • Step C Preparation of (2S,2'S) 2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N-[N ⁇ -(4- methylbenzenesulfonyl)-S-tryptophanyl]-2,6-diaminohexanal
  • the title compound was prepared from the previous derivative (400 mg, 0.59 mmol) by oxidation under Swern's reaction conditions using oxalyl chloride (110 ⁇ L, 1.2 mmol), DMSO (166 ⁇ L, 2.4 mmol), and triethylamine (2 mL) (Organic Syntheses, Collective Volume VII, p. 258-263).
  • the crude material was purified by flash chromatography using a solyent gradient from 39.T to 19:1 CH 2 Cl 2 /MeOH. The product was isolated as a yellowish solid (330 mg, 83% yield).
  • Step B Preparation of (2S,2'S) 2-N-isobutyl-2-N-(4-methylberrzenesulfonyl)-6-N-[N ⁇ -(4- methoxybenzenesulfonyl)-S-phenylalanyl]-2,6-diaminohexanol
  • Step B Preparation of N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-Ne-[N' ⁇ -(4- methoxyphenylacetyl)-L-phenylalanyl]-L-lysine
  • the title compound was prepared from N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-L-lysine hydrochloride (100 mg, 0.25 mmol, example 1, step E) as described in general procedure Be. using N ⁇ -(4-methoxyphenylacetyl)-L-phenylalanine (80 mg, 0.25 mmol) which was prepared in step A of this example. The final product was triturated with ether to yield 85 mg (18%) of the desired material.
  • Step B Preparation of ⁇ ? ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-Ne-(N -dihydrocinnamoyl- L-tryptophanyl)-L-Iysine
  • the title compound was prepared from N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-L-lysine hydrochloride (400 mg, 1.0 mmol, example 1 , step E) as described in general procedure Be using N ⁇ -dihydrocinnamoyl-L-tryptophan (360 mg, 1.0 mmol) which was prepared in step A of this example.
  • the final product was triturated with ether to yield 612 mg (85% pure) of the crude material. Purification of 100 mg of the crude material by HPLC gave 45 mg (45%) of pure adduct.
  • LC-MS 641.5 (M + H) + , 99% pure. " ' .
  • Step A Preparation of N ⁇ -isobutyl- ⁇ -(4-nitrobenzenesulfonyl)-Ne-benzyloxycarbonyl-L-lysine methyl ester
  • Step B Preparation of N ⁇ -(4-aminobenzenesulfonyl)-N ⁇ -isobutyl-L-lysine methyl ester
  • step A The title compound was obtained by catalytic hydrogenation of N ⁇ -isobutyl-N ⁇ -(4- nitrobenzenesulfonyl)-Ne-benzyloxycarbonyl-L-lysine methyl ester (step A) as described for the preparation of N ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-L-lysine methyl ester (example 65, step C). This compound was used without purification in the next step.
  • Step C Preparation of N ⁇ -(4-aminobenzenesulfonyl)-N ⁇ -isobutyl-Ne-[N' ⁇ -(4- methylbenzenesulfonyl)-L-tryptophanyl]-L-lysme methyl ester
  • the title compound was prepared in the same manner than example 69 using N ⁇ -(4- arninobenzenesulfonyl)-Ni ⁇ -isobuty]-L -lysine methyl ester (step B) as the starting material.
  • the yield of this reaction was 65%.
  • Step D Preparation of (25,2'S) 2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-6-N-[ ⁇ p ⁇ -(4- methylbenzenesulfonyl)-5-tryptophanyl]-2,6-diaminohexanol N ⁇ -(4-am
  • 2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-6-N-[ ⁇ p ⁇ -(4- methylbenzenesulfonyl)-5-tryptophanyl]-2,6-diaminohexanol N ⁇ -(4-am devisizenesulfonyl)-N ⁇ -isobutyl-Ne-[N ⁇ -(4 ⁇
  • step C L-lysine methyl ester (step C) was reduced with LiAlH 4 following the indications of general procedure G. The final product was obtained in 37% yield.
  • Step A Preparation of N ⁇ -isobutyl-N ⁇ -(4-methoxybenzenesulfonyl)-N6-benzyloxycarbonyl-L- lysine methyl ester
  • Step B Preparation of N ⁇ -isobutyl-N ⁇ -(4-methoxybenzenesulfonyl)-L-lysine methyl ester
  • step A The title compound was obtained quantitatively by catalytic hy drogenation of N ⁇ -isobutyl-N ⁇ -(4- methoxybenzenesulfonyl)-Ne-benzyloxycarbonyI-L-lysine methyl ester (step A) as described for the preparation of ⁇ -isobutyl-N ⁇ -(4-methylbenzenesulfonyl)-L-lysine methyl ester (example 65, step C). This compound was used without purification in the next step.
  • Step C Preparation . of N ⁇ -isobutyLN ⁇ -(4-methoxybenzenesulfonyl)-Ne-(N' ⁇ -tert- butoxycarbonyl-L-tryptophanyl)-L-lysine methyl ester
  • the title compound 1 was prepared in the same manner " as in the example 65 (step D) using N ⁇ - isobutyl-N ⁇ -(4-methoxybenzenesulfonyl)-L-lysine methyl ester (step B) and commercially available N ⁇ -te7Y-butoxycarbonyl-L-tryptophan as the starting materials. The yield of this reaction was 86%.
  • Step D Preparation of (25,2'S) 2-N-isobutyl-2-N-(4-methoxybenzenesulfonyl)-6-N-(N' ⁇ -tgrt- butoxycarbonyl-S-tryptophanyl)-2,6-diaminohexanol N ⁇ -isobutyl-N ⁇ -(4-methoxybenzenesulfonyl)-Ne-(N , ⁇ -tert-butoxycarbonyl-L-tryptophanyl)-L- lysine methyl ester (step C) was reduced with LiAlH 4 following the indications, of general procedure G. The final product was obtained in 81% yield.

Abstract

A compound selected from the group consisting of a compound of formula I or formula II: (I) (II) wherein n is 3 or 4 wherein Cx is selected from the group consisting of -COOM, COOR6, -CHO, -CH2OR7, -CH2OCOR8, -CONHR9 and -CONR10R11, wherein M is an alkali metal or alkaline earth metal, wherein R2 is selected from the group consisting of a benzenesulfonyl group of formula III, or formula IV : (III) (IV) and when the compound of formula (I) and (II) comprises an amino group pharmaceutically acceptable ammonium salts thereof, wherein R1, R2, Cx, n, R3, R4, R5, Y are as defined in the specification.

Description

HIV PROTEASE INHIBITORS BASED ON AMINO ACID DERIVATIVES
TECHNICAL FIELD OF THE INVENTION
The present invention concerns amino acid derivatives possessing aspartyl protease inhibitory properties, in particular Ne-amino acid substituted L-lysine derivatives (and analogs) possessing aspartyl protease inhibitory properties. It describes also the synthetic methodology used to make these derivatives and their biological applications. In addition, this invention relates to different pharmaceutical compositions comprising these compounds. The compounds and the pharmaceutical compositions of this invention have been shown to inhibit the activity of HIN aspartyl protease, an enzyme essential for virus maturation. The inhibitory property may be advantageously used to provide compounds with antiviral properties against HIN viruses, including the HIN-1 and HIV-2 viruses.
BACKGROUND OF THE INVENTION
The HIN (human immunodeficiency virus) retrovirus is responsible for causing the disease Icnown as AIDS (acquired immunodeficiency syndrome). HIN infection is characterized by a period immediately following infection, called asymptomatic, which is devoid- of clinical manifestations in the patient. Progressive HIN-induced destruction of the immune system then leads to increased susceptibility to opportunistic infections, which eventually produces a syndrome called AIDS-related complex (ARC) characterized by symptoms such as persistent generalized lymphadenopathy, fever, weight loss, followed itself by full blown AIDS.
As the first step of its replication cycle, the HIN-1 retrovirus attaches primarily to the CD4 receptor (a 58 kDa transmembrane protein) to gain entry into susceptible cells, through high- affinity interactions between the viral envelope glycoprotein (gp 120) and a specific region of the CD4 molecule found in CD4 (+) T-helper lymphocytes and other cells carrying the receptor (Lasky L.A. et al., Cell vol. 50, p. 975-985 (1987)). The HIN genetic material, in the form of RΝA. is then transcribed into DΝA by a viral enzyme carried by the virus called reverse transcriptase. The viral DΝA now called pro virus is then transported into the cell nucleus in the form of a preintegration complex and' attached to the cell DΝA by another viral enzyme called integrase. Following integration, the viral DΝA then serves as a template for viral gene expression by the host transcription system. The primary RΝA transcripts made from the provirus are synthesized by the host cell RΝA.polymerase II whose activity is modulated by two virus-encoded proteins, Tat and Rev. The viral proteins are expressed mainly in the form of polyproteins. After the infected cell has produced all the different HIV polyproteins and genetic material, they assemble at the cell membrane and are released from the cell in the form of an immature viral particle. A third viral enzyme known as protease then cleaves the polyproteins to give the mature, infectious viral particle. The polyproteins that are cleaved by the HIV protease are the Gag and Gag-Pol precursors, whose cleavage gives rise to structural proteins and viral enzymes.
A number of synthetic antiviral agents have been designed to block various stages in the replication cycle of HIV. although only those developed against the viral enzymes have reached the market so far. The latter include compounds which block viral reverse transcriptase (for example, didanosine and zidovudine (AZT))5 or the viral protease (for example, ritonavir and indinavir). Although these drugs have improved significantly the survival time and quality of life of AIDS patients, the administration of most of these agents leads to unwanted side effects, such as anemia, neurotoxicity and bone marrow suppression.
Anti-protease compounds represent the most-recent drugs developed to block HIV replication. These compounds inhibit the formation of infectious virions by interfering with the processing of viral polyprotein precursors by the viral protease. The antiviral potential of HIV protease inhibitors was first demonstrated using peptidic inhibitors. Such peptidic compounds, however, are typically large and complex molecules that tend to exhibit poor bioavailability and limited stability in the body. New compounds devoid of these drawbacks are urgently needed to treat HIV infections. In addition, mutations arising during HIV replication lead to resistance to the currently available protease inhibitors, so new compounds with original structure are desirable to fight these resistant viral strains.
SUMMARY OF THE INVENTION
The present invention provides a novel class of compounds, including their pharmaceutically acceptable derivatives. These compounds have an affinity for aspartyl proteases, in particular, HIV aspartyl protease. Therefore, these compounds are useful as inhibitors of such proteases. These compounds can be used alone or in combination with other therapeutic or prophylactic agents for the treatment or prophylaxis of viral infection.
According to a preferred embodiment, the compounds of this invention are capable of inhibiting HIV viral replication in human CD4+ T-cells, by inhibiting the ability of HIV aspartyl protease to catalyse the hydrolysis of peptide bonds present in viral Gag and Gag-Pol polyproteins. These novel compounds can thus serve to reduce the production of infectious virions from acutely and chronically infected cells, and can inhibit the initial or further infection of host cells. Accordingly, these compounds are useful as therapeutic and prophylactic agents to treat or prevent infection by HIV-1 and HIV-2, which may result in asymptomatic infection, AIDS- related complex (ARC), acquired immunodeficiency syndrome (AIDS), AIDS-related dementia, or similar diseases of the immune system, and related viruses such as HTLV-I and HTLV-II, and simian immunodeficiency virus.
It is the main objective of this invention to provide a novel class of molecules that are aspartyl protease inhibitors, and particularly, HIN aspartyl protease inhibitors.
The present invention relates to a class of Ne-amino acid substituted L-lysine derivatives (including its lower and higher homologues and analogs) as well as their pharmaceutically acceptable derivatives (e.g. salts). Accordingly, the present invention in accordance with one aspect thereof provides a compound of formula I
Figure imgf000005_0001
a compound of formula II
Figure imgf000005_0002
II
and when the compound of formula I and II comprises an amino group pharmaceutically acceptable ammonium salts thereof,
wherein n is 3 or 4
wherein Y is 0, S or N-CN
wherein Cx is selected from the group consisting of -COOM, COOR^, -CHO, -CH2OR7, -
Figure imgf000005_0003
-CONHRg and -CONR,0R„, wherein M is an alkali metal (e.g. Na, K, Cs, etc) or alkaline earth metal (Ca, Mg, etc.), wherein R, is selected from the group consisting of a straight or branched alkyl group of 1 to 6 carbon atoms, a cycloalkylalkyl group having 3 to 6 carbon atoms in the cycloalkyl part thereof and 1 to 3 carbon atoms in the alkyl part thereof,
wherein R-, is selected from the group consisting of a benzenesulfonyl group of formula III,
Figure imgf000006_0001
and a thiophenesulfonyl group of formula IV,
Figure imgf000006_0002
IV
wherein R- is selected from the group consisting of H, a straight or branched alkyl group of 1 to 6 carbon atoms, a phenyl or a benzyl group
wherein R., is selected from the group consisting of H, a group of formula Ilia
Figure imgf000006_0003
Ilia a group of formula IVa
Figure imgf000007_0001
IVa
C6H, ,-, C5H10N-CH2CH2-, OC4H8N-CH2CH2- (i.e. morpholine- 4-CH2CH2-), C6H5CH2CH2-, 2,3- (CH30)2C6H3CH2-, C6H5-, 2-C5H4N (i.e.2-pyridinyl), 3-C5H4N (i.e.3-pyridinyl), 4-C5H,N (i.e. 4-pyridinyl), 3-quinolyl, C6H5CS-, 2-naphthyl-SO2- and a group of formula R4C-CO-, R4C being selected from the group consisting of a straight or branched alkyl group of 1 to 6 carbon atoms, a cycloalkylalkyl group having 3 to 6 carbon atoms in the cycloalkyl part thereof and 1 to 3 carbon atoms in the alkyl part thereof, (e.g. CH3-, wo-butyl, wo-propyl, tert-butyl, tert-butyl- CH:-), CF3, 1 -pyrrolidinyl, 4-morpholinyl>tetrahydro-3-furanyloxy,4-CH3OC6H4NH-5 CH3NH-, HOCH2CH2NH-, 9-fluorenyl-CH20-, rert-butylO-, iso-butylO-, C6H5CH2O-, CH3O-, unsubstituted C6H5-, C6H5- substituted by one or more members (e.g. one or two) selected from the group consisting of F, Cl, Br, I, -CF3, -NO2, -NRι0Rn, -NHCOR10. -OR10, -SR10, -COOR10, -COR,o and -CH2OH, unsubstituted C6H5CH2-, C6H5CH2- substituted by one or more members (e.g. one or two) selected from the group consisting of F, Cl, Br, I, -CF3, -NO2, -NR10RU, - NHCOR10, -OR10; -SR10, -COOR10, -COR10 and -CH2OH, unsubstituted C6H5CH2CH2-, and C6H5CH2CH2- substituted by one or more members (e.g. one or two) selected from the group consisting of F, CL Br, I, -CF3, -NO2, -NR,0R„, -NHCOR10, -OR10, -SR10, -COOR10, -COR10 and -CH,OH,
wherein R5 is selected from the group consisting of H, a straight or branched alkyl group of 1 to 8 carbon atoms, (e.g. CH3-, CH3CH2CH2-5 CH3CH2CH2CH2-5 iso-C hl9-, C6H„CH2-), HOCH,-, C6H5CH2OCH2-, benzyl-OCH(CH3), HO2CCH2-, HO2CCH2CH2> NC-CH2-, H2NC(O)CH,-, H,NC(0)CH2CH2-, 4-CH3C6H4CH2SCH2-, CH3SCH2CH2-, H2NCH2CH2CH2CH2-, C6H5-, C6HsCH2-; C6H5CH(OH)-, C6H5CH(CN)-, C6F5CH2-, 4-(9-fluorenylmethoxycarbonyl)-NHCH2- CόH,CH2-5 C5H4N-2-CH2- (i.e. pyridine-2-CH2-), C5H4N-3-CH2- (i.e. pyridine-3-CH2-), C5H4N- 4-CH2- (i.e. pyridine-4-CH2-). 2-thiophene-CH2-_ indole-3-CH2-, 2-benzothiophene-CH2-, Nτ- benzyl-imidazole-4-CH2- , imidazole-4-CH2-, thiazole-4-CH2- and substituted benzyl (e.g. benzyl substituted by a group being as defined for R]2 below, e.g. 4-tert-butyl-C6H4CH2-, 4- HOC6H4CH2-, 4-benzyl-O-C6H4CH2-5 4-NO2C6H4CH2-, 2-FC6H4CH2-, 3-FC6H4CH2-, 4- FC6H4CH2-),
wherein Ra represents a member selected from the group consisting of
Figure imgf000008_0001
wherein Met is a methylene linked to the α' nitrogen
wherein R6 is selected from the group consisting of H, a straight or branched alkyl group of 1 to 4 carbon atoms and glycyl
wherein R7 is selected from the group consisting of H, a straight or branched alkyl group of 1 to 4 carbon atoms
wherein Rg is selected from the group consisting of a straight or branched alkyl group of 1 to 6 carbon atoms, a cycloalkyl group having 3 to 6 carbon atoms, a cycloalkylalkyl group having 3 to 6 carbon atoms in the cycloalkyl part and 1 to 3 carbon atoms in the alkyl part thereof,
wherein R9 is selected from the group consisting of H, a straight or branched alkyl group of 1 to 4 carbon atoms, -OH, -NH2 and -CH2CH2OH
wherein R10 and Rπ are independently selected from the group consisting of H, a straight or branched alkyl group of 1 to 4 carbon atoms wherein m is 0 or 1
wherein o is 0 or 1
wherein R,2 and R)3 are independently selected from the group consisting of H, a straight or branched alkyl group of 1 to 4 carbon atoms, F, Cl, Br, I, -CF3, -NO2, -NR,0Rπ, -NHCOR10, ,- OR,05 -OCH,C6H5, -SR]0, -COOR,0, -COR]0 and -CH2OH, R,0 and Rn being as defined herein.
More particularly, this invention provides a compound of formula IA
Figure imgf000009_0001
IA
and when the compound of formula I A comprises an amino group pharmaceutically acceptable ammonium salts thereof,
wherein n is 3 or 4
wherein Y is O, S or N-CN
wherein Cx is selected from the group consisting of -COOM, COOR^,. -CHO, -CH2OR7, - CH2OCORs, -CONHRp and -CONR10R,„ wherein M is an alkali metal (e.g. Na, K, Cs, etc) or alkaline earth metal,
wherein R, is selected from the group consisting of a straight or branched alkyl group of 1 to 6 carbon atoms, a cycloalkylalkyl group having 3 to 6 carbon atoms in the cycloalkyl part thereof and 1 to 3 carbon atoms in the alkyl part thereof, wherein R2 is selected from the group consisting of a benzenesulfonyl group of formula III,
Figure imgf000010_0001
III
and a thiophenesulfonyl group of formula IV,
Figure imgf000010_0002
IV
wherein R3 is selected from the group consisting of H, a straight or branched alkyl group of 1 to 6 carbon atoms, a phenyl or a benzyl group
wherein R4 is selected from the group consisting of H, C6Hn-5 C5H10N-CH2CH2-5 OC4H8N- CH2CH2- (i.e. morpholine- 4-CH2CH2-); C6H5CH2CH2-, 2,3-(CH3O)2C6H3CH2-, C6H5-, 2-C5H4N, 3-C5H4N, 4-C5H4N, 3-quinolyl, CH3CO-, CF3CO, C6H5CO-5 C6H5CS-, 4-CH3OC6H4CH2CO-, C6H5CH2CH2CO-; iso-butyl-CO-, wo-propyl-CO-, te;-t-butyl-CO-, tert-butyl-CH2CO-, 1- pyrrolidine-CO-, 4-morpholine-CO-, carbotetrahydro-3-furanyloxy, 4-CH3OC6H4NHCO-, CH,NHCO-, HOCH2CH2NHCO-, 9-fluorenylmethoxy carbonyl, tert-butylO-CO-, zso-butylO-CO- , C6H5CH20-CO-, CH3O-CO-, C6H5SO2-54-CH3C6H4S02-54-CF3C6H4SO2-, 4-NO2C6H4SO2-, 4- NH,C6H4SO2-, 4-AcNHC6H4SO2-, 4-FC6H4SO2-, 4-ClC6H4SO2-, 4-BrC6H4SO2-, 4- CH3OC6H4SO2-, 2-thiophene-SO2- and 2-naphthyl-SO2-
wherein R5 is selected from the group consisting of H, CH3-, CH3CH2CH2-,CH3CH2CH2CH2-. wo-C4H9-, C6HnCH2-, HOCH2-, C6H5CH2OCHr benzyl-OCH(CH3), HO2CCH2-5 H02CCH,CH,-, NC-CH2-, H2NC(O)CH2-, H2NC(O)CH2CH2-, 4-CH3C6H4CH2SCH2-, CH3SCH,CH2- H2NCH2CH2CH2CH2-3 C6H5-, C6H5CH2-, C6F5CH2-, 4-tert-butyl-C6H4CH2-s 4- HOC6H4CH2-, 4-berτzyl-O-C6H4CH2-, 4-NO2C6H4CH2-, 4-(9-fluorenylmethoxycarbonyl)NHCH2- C6H4CH2-, 2-FC6H4CH2-, 3-FC6H4CH2-, 4-FC6H4CH2-, C5H4N-2-CH2-5 C5H4N-3-CH2-, C5H4N-4- CH2-, 2-thiophene-CH2-ι indole-3-CH2-, 2-benzothiophene-CH2-, Nτ-benzyl-imidazole-4-CH2- and thiazole-4-CH2-
wherein F^ is selected from the group consisting of H, a straight or branched alkyl group of 1 to 4 carbon atoms and glycyl
wherein R7 is selected from the group consisting of H, a straight or branched alkyl group of 1 to 4 carbon atoms
wherein R8 is selected from the group consisting of a straight or branched alkyl group of 1 to 6 carbon atoms, a cycloalkyl group having 3 to 6 carbon atoms, a cycloalkylalkyl group having 3 to 6 carbon atoms in the cycloalkyl part and 1 to 3 carbon atoms in the alkyl part thereof,
wherein R-j is selected from the group consisting of H, a straight or branched alkyl group of 1 to 4 carbon atoms, -OH, -ΝH2 and -CH2CH2OH
wherein R,0 and R,, are independently selected from the group consisting of H, a straight or branched alkyl group of 1 to 4 carbon atoms .
wherein m is 0 or 1
wherein o is 0 or 1
wherein R12 and R13 are independently selected from the group consisting of H, a straight or branched alkyl group of 1 to 4 carbon atoms, F, Cl, Br, I, -CF3, -NO2, -NR10Rπ, -NHCOR10, - OR10, -OCH2C6H5: -SR10, -COOR10, -COR10 and -CH2OH, R10 and R„ being as defined herein.
This invention also provides a compound of formula la
Figure imgf000012_0001
and when the compound of formula la comprises an amino group pharmaceutically acceptable ammonium salts thereof,
wherein Cx is selected from the group consisting of -COOM and -CH2OH, M being an alkali metal or alkaline earth metal,
wherein R1A is selected from the group consisting of a straight or branched alkyl group of 1 to 6 carbon atoms, a cycloalkylalkyl group having 3 to 6 carbon atoms in the cycloalkyl part thereof and 1 to 3 carbon atoms in the alkyl part thereof,
wherein R2A and R4A are independently selected from the group consisting of H, a straight or branched alkyl group of 1 to 4 carbon atoms, F, Cl, Br, I, -CF3, -NO2, -NR]0R,,, -NHCOR]0, - OR,0, -OCH,C6H5J -SR10, -COOR10, -COR10 and -CH2OH,
wherein R is selected from the group consisting of H, a straight or branched alkyl group of 1 to 8 carbon atoms, (e.g. CH3-, CH3CH2CH2-,CH3CH2CH2CH2-, /.rø-C4H9-, C6H, ,CH2-)5 HOCH,-, C6H5CH2OCH2-, benzyl -OCH(CH3), HO2CCH2-, H02CCH2CH2-, NC-CH2-5 H2NC(O)CH2-, H2NC(0)CH2CH2-. 4-CH3C6H4CH2SCH2-, CH3SCH2CH ι H2NCH2CH2CH2CH2-, C6H5-, C6H5CH2-, C6H5CH(OH)-; C6H5CH(CN)-, C6F5CH2-,4-(9-fluorenylmethoxycarbonyl)-NHCH2- C6H4CH2-, C5H4N-2-CH2- (i.e. pyridine-2-CH2-), C5H4N-3-CH2- (i.e. pyridine-3-CH2-), C5H4N- 4-CH2- (i.e. pyridine-4-CH2-), 2-thiophene-CH2-> indole-3-CH2-, 2-benzothiophene-CH2-, Nτ- benzyl-imidazole-4-CH2-, imidazole-4-CH2-,thiazole-4-CH2-and substituted benzyl (e.g. benzyl substituted by a group as defined for R2A, e.g.4-ter/-butyl-C6H4CH2-54-HOC6H4CH2-, 4-benzyl- O-C6H4CH2-, 4-ΝO2C6H4CH2-, 2-FC6H4CH2-, 3-FC6H4CH2-54-FC6H4CH2-)5 and wherein R10 and Rn are as defined herein.
This invention also provides a compound of formula lb
Figure imgf000013_0001
and when the compound of formula lb comprises an amino group pharmaceutically acceptable ammonium salts thereof,
wherein Cx is selected from the group consisting of -COOM, and -CH2OH, M being an alkali metal or alkaline earth metal,
wherein R1A is selected from the group consisting of a straight or branched alkyl group of 1 to 6 carbon atoms, a cycloalkylalkyl group having 3 to 6 carbon atoms in the cycloalkyl part thereof and 1 to 3 carbon atoms in the alkyl part thereof,
wherein R2A and R4B are independently selected from the group consisting of H, a straight or branched alkyl group of 1 to 4 carbon atoms, F, Cl, Br, I, -CF3, -NO2, -NR10Rn, -NHCOR,0, - OR)05 -OCH2C6H5, -SR10, -COOR10, -COR10 and -CH2OH,
wherein R5A is selected from the group consisting of H, a straight or branched alkyl group of 1 to 8 carbon atoms, (e.g. CH3-, CH3CH2CH2-,CH3CH2CH2CH2-5 iso-C U9-, C6HnCH2-), HOCH2-, C6H5CH2OCH , benzyl-OCH(CH3), HO2CCH2-, HO2CCH2CH2-, NC-CH2-, H2NC(O)CH2-, H2NC(O)CH2CH2-, 4-CH3C6H4CH2SCH2-,
Figure imgf000013_0002
H2NCH2CH2CH2CH2-, . C6H5-, C6HSCH,-, C6H5CH(OH)-, C6H5CH(CN)-, C6F5CH2-,4-(9-fluorenylmethoxycarbonyl)-NHCH2- C6H4CHr, C5H4N-2-CH2- (i.e. pyridine-2-CH2-), C5H4N-3-CH2- (i.e. pyridine-3-CH2-), C5H4N- 4-CH2- (i.e. pyridme-4-CH,-), 2-thiophene-CH2- indole-3-CH2-, 2-benzothiophene-CH2-, Nτ- benzyl -imidazoIe-4-CH2-, imidazole-4-CH2-,thiazole-4-CH2- and substituted benzyl (e.g. benzyl substituted by a group as defined for R2A, e.g.4-tert-butyl-C6H4CH2-, 4-HOC6H4CH2-, 4-benzyl- O-C6H4CH2-, 4-NO2C6H4CH2-, 2-FC6H4CH2-, 3-FC6H4CH2-, 4-FC6H4CH2-),
and wherein R10 and Ru are as defined herein.
This invention also provides a compound of formula Ic
Figure imgf000014_0001
and when the compound of formula Ic comprises an amino group pharmaceutically acceptable ammonium salts thereof,
wherein Cx is selected from the group consisting of -COOM and -CH2OH, M being an alkali metal or alkaline earth metal,
wherein RIA is selected from the group consisting of a straight or branched alkyl group of 1 to 6 carbon atoms, a cycloalkylalkyl group having 3 to 6 carbon atoms in the cycloalkyl part thereof and 1 to 3 carbon atoms in the alkyl part thereof,
wherein R2B and R4B are independently selected from the group consisting of H, a straight or branched alkyl group of 1 to 4 carbon atoms, F, Cl, Br, I, -CF3, -NO2, -NR10R,), -NHCOR)0, - ORI0, -OCH2C6H5, -SR,0, -COOR)0, -COR10 and -CH2OH,
wherein RJA is selected from the group consisting of H, a straight or branched alkyl group of 1 to 8 carbon atoms, (e.g. CH3-, CH3CH2CH2-,CH3CH2CH2CH2-, zso-C4H9-, C6HUCH2-), HOCH2-, C6H5CH2OCH2-, benzyl-OCH(CH3), HO2CCH2-, HO2CCH2CH,-, NC-CH,-, H2NC(O)CH2-. H2NC(0)CH2CH2-, 4-CH3C6H4CH2SCH2-, CH3SCH2CH > H2NCH2CH2CH2CH2-, C6H5-, C6H5CH2-, C6H5CH(OH)-, C6H5CH(CN)-, C6F5CH2-, 4-(9-fluorenylmethoxycarbonyl)-NHCH2- C6H4CH2-, C5H4N-2-CH2- (i.e. pyridine-2-CH2-), C5H4N-3-CH2- (i.e. pyridine-3-CH2-), C5H4N- 4-CH2- (i.e. pyridiιιe-4-CH2-)s 2-thiophene-CH2- indole-3-CH2-, 2-benzothiophene-CH2-, Nτ- benzyl-imidazole-4-CH2-, imidazole-4-CH2-, thiazole-4-CH2- and substituted benzyl (e.g. benzyl substituted by a group as defined for R2A, e.g.4-tert-butyl-C6H4CH2-, 4-HOC6H4CH2-, 4-benzyl- O-C6H4CH2-, 4-ΝO2C6H4CH2-, 2-FC6H4CH2-, 3-FC6H4CH2-, 4-FC6H4CH2-),
and wherein R,nO a • nd R, , are as defined herein.
This invention also provides a compound of formula Id
Figure imgf000015_0001
and when the compound of formula Id comprises an amino group pharmaceutically acceptable ammonium salts thereof,
wherein Cx is selected from the group consisting of -COOM and -CH2OH, M being an alkali metal or alkaline earth metal,
wherein R]A is selected from the group consisting of a straight or branched alkyl group of 1 to 6 carbon atoms, a cycloalkylalkyl group having 3 to 6 carbon atoms in the cycloalkyl part thereof and 1 to 3 carbon atoms in the alkyl part thereof, wherein R4A is selected from the group consisting of H. a straight or branched alkyl group of 1 to 4 carbon atoms, F, Cl, Br, I, -CF3, -NO2, -NR^R,,, -NHCOR10, -OR10, -OCH2C6H5, -SRlθ5 - COOR10, -COR10 and -CH2OH,
wherein R4C is selected from the group consisting of a straight or branched alkyl group of 1 to 6 carbon atoms, a cycloalkylalkyl group having 3 to 6 carbon atoms in the cycloalkyl part thereof and 1 to 3 carbon atoms in the alkyl part thereof, (e.g. CH3-, iso-butyl, z'so-propyl, tert-butyl, tert- butyl-CH2-), CF3, pyirolidine, 4-morpholine, tetrahydro-3-furanyloxy, 4-CH3OC6H4NH-, CH3NH-, HOCH2CH2NH-, 9-fluorenyl-CH2O-5 tert-butylO-, zso-butylO-, C6H5CH2O-, CH3O-, unsubstituted C5H5-5 C6H5- substituted by one or more members (e.g. one or two) selected from the group consisting of F, Cl, Br, I, -CF3, -NO2, -NR,0Rπ, -NHCOR10, -OR!0, -SR10, -COOR10, -COR10 and -CH2OH, unsubstituted C6H5CH2-, C6H5CH2- substituted by one or more members (e.g. one or two) selected from the group consisting of F, Cl, Br, I, -CF3, -N02, -NR,0Rπ, - NHCOR10, -OR)0, -SR]0, -COORi0, -CORJ0 and -CH2OH, unsubstituted C6H5CH2CH2-, and C6H5CH2CH2- substituted by one or more members (e.g. one or two) selected from the group consisting of F, Cl, Br, I, -CF3, -NO2, -NR,0R,„ -NHCOR10, -OR10, -SR10, -COOR1Q, -COR10 and -CH2OH,
wherein R5A is selected from the group consisting of H, a straight or branched alkyl group of 1 to 8 carbon atoms, (e.g. CH3-, CH3CH2CH2-,CH3CH2CH2CH2-, zso-C4H9-, C6HUCH2-), HOCH , C6H5CH2OCH2-5 benzyl-OCH(CH3), HO2CCH2-, HO2CCH2CH2-, NC-CH2-, H2NC(O)CH2-, H2NC(0)CH2CH2-, 4-CH3C6H4CH2SCH2-, CH3SCH2CH2-, H2NCH2CH2CH2CH2-, C6H5-, C6H 5CH2-, C6H5CH(OH)-, C6H5CH(CN)-, C6F5CH2-54-(9-fluorenyImethoxycarbonyl)-NHCH2- C6H4CH2-, C5H4N-2-CH2- (i.e. pyridine-2-CH2-), C5H4N-3-CH2- (i.e. pyridine-3-CH2-), C5H4N- 4-CH2- (i.e. pyridine-4-CH2-), 2-thiophene-CH2-> indole-3-CH2-, 2-benzothiophene-CH2-, Nτ- benzyi-imidazole-4-CH2- , imidazole-4-CH2-5 thiazole-4-CH2- and substituted benzyl (e.g. benzyl substituted by a group as defined for R2A, e.g.4-tert-butyl-C6H4CH2-, 4-HOC6H4CH2-, 4- benzyl-O-QH4CH2-, 4-Ν02C6H4CH,-, 2-FC6H4CH2-, 3-FC6H4CH2-, 4-FC6H4CH2-),
and wherein R,„ and R,, are as defined herein. In addition, this invention provides pharmaceutical compositions in which these novel compounds of formula I or II, (as well as of formula IA, la, lb, Ic and Id) derived from L-lysine or L-lysine derivatives (as well as lower and higher homologues) are used to inhibit aspartyl proteases, including HIV aspartyl protease, thus providing protection against HIV infection.
Thus the present invention provides a pharmaceutical composition comprising a pharmaceutically acceptable carrier and a pharmaceutically effective amount of at least one compound of formula I, formula II (as well as of formulae IA, la, lb, Ic, and Id) and as applicable pharmaceutically acceptable ammonium salts thereof.
The terms "HIV protease" and "HIV aspartyl protease" are used interchangeably and refer to the aspartyl protease encoded by the human immunodeficiency virus type 1 or 2. In a preferred embodiment of this invention, these terms refer to the human immunodeficiency virus type 1 aspartyl protease.
The term "pharmaceutically effective amount" refers to an amount effective in treating HIV infection in a patient.
The term "prophylactically effective amount" refers to an amount effective in preventing HIV infection in a patient. As used herein, the term "patient" refers to a mammal, including a human.
The terms "pharmaceutically acceptable carrier", "pharmaceutically acceptable adjuvant" and "physiologically acceptable vehicle" refer to a non-toxic carrier or adjuvant that may be administered to a patient, together with a compound of this invention, and which does not destroy the pharmacological activity7 thereof.
The compounds of this invention include pharmaceutically acceptable derivatives of the compounds of formula I, formula II (as well as of formulae IA, la, lb, Ic, and Id) and as applicable pharmaceutically acceptable ammonium salts thereof. A "pharmaceutically acceptable derivative" means any pharmaceutically acceptable salt, ester, or salt of such ester, of a compound of this invention or any other compound which, upon administration to a recipient, is capable of providing (directly or indirectly) a compound of this invention or an antivirally active metabolite or residue thereof.
Salts derived from appropriate bases include alkali metal (e.g., sodium), alkaline earth metal (e.g., magnesium), ammonium and N - (CMaIkyi)4 + salts.
The compounds of this invention contain one or more asymmetric carbon atoms and thus may occur as racemates and racemic mixtures, single enantiomer, diastereomeric mixtures and individual diastereoisomers. All such isomeric forms of these compounds are expressly included in the present invention. Each stereogenic carbon may be of the R or S configuration.
Combinations of substituents and variables envisioned by this invention are only those that result in the formation of stable compounds. The term "stable", as used herein, refers to compounds which possess stability sufficient to allow manufacture and administration to a mammal by methods known in the art. Typically, such compounds are stable at a temperature of 40°C or less, in the absence of moisture or other chemically reactive conditions, for at least a week.
Pharmaceutically acceptable salts of the compounds of this invention include those derived from pharmaceutically acceptable inorganic and organic acids and bases. Examples of such acid salts include: acetate, adipate, alginate, aspartate, benzoate, benzenesulfonate, bisulfate, butyratc, citrate, camphorate, camphorsulfonate, cyclopentanepropionate, digluconate, dodecylhydrogensulfate, dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptanoate, glycerophosphate, glycollate, hemisulfate, heptanoate, hexanoate, hydrochloride, hydrobromide, hydroiodide, 2-hydroxyethanesulfonate, lactate, maleate, malonate, methanesulfonate, 2- naphthylsulfonate, nicotinate, nitrate, oxalate, pamoate, pectinate, perchlorate, persulfate, 3- phenylpropionate. phosphate, picrate, pivalate, propionate, salicylate, succinate, sulfate, tartrate, thiocyanate, tosylate, and undecanoate.
This invention also envisions the quaternization of any basic nitrogen containing groups of the compounds disclosed herein. The basic nitrogen can be quateraized with any agents known to those of ordinary skill in the art including, for example, lower alkyl halides, such as methyl, ethyl, propyl and butyl chlorides, bromides and iodides; dialkyl sulfates including dimethyl, diethyl, dibutyl and diamyl sulfates; long chain halides such as decyl, lauryl, myristyl and stearyl chlorides, bromides and iodides, and aralkyl halides including benzyl and phenethyl bromides. Water or oil-soluble or dispersible products may be obtained by such quaternization.
The compounds of this invention are readily prepared using conventional techniques from commercially available and inexpensive starting materials. The relative ease of synthesis of the products described herein represents a marked advantage for the large scale preparation of these compounds. In general, the derivatives of the present invention may be readily obtained through sequences recognized by those knowledgeable in the art as straightforward. These sequences are presented in schemes 1 to 7 discussed below.
Scheme 1 illustrates a generic example for the preparation of a key intermediate needed for the synthesis of HIV protease inhibitors.
Note: a) For scheme 1 , R, represents an alkyl or cycloalkylalkyl side chain as defined above b) R2 represents a benzenesulfonyl group of formula III, a thiophenesulfonyl group of formula IV. a 1 -naphthylsulfonyl, a 2-naphthylsulfonyl or a 8-quinolinesulfonyl group as defined above
As shown in scheme 1, the Nα,Nα-disubstituted L-lysine derivative 5 was obtained from commercially available L-lysine 1 in a four-step reaction sequence. This preparation uses the cyclic form of L-lysine in order to manipulate the N -amino group without the need for protective groups. First, L-lysine was transformed into L- -amino-e-caprolactam 2 upon treatment with hydrochloric acid in methanol followed by neutralization with sodium hydroxide. The caprolactam 2 is also commercially available. Reductive alkylatibn of derivative 2 with an appropriate aldehyde and ΝaBH(OAc)3 in dichloroethane lead to the Nα-alkylamirio-e- caprolactam 3. Then, sulfonation with an arylsulfonyl chloride or a substituted-arylsulfonyl chloride in the presence of triethylamine in dichloromethane gave compound 4 in excellent yields. The N ,Nα-disubstituted L-lysine derivative 5 was obtained quantitatively by acid hydrolysis of the cyclic amide 4. Scheme 1
Figure imgf000020_0001
1 , L-lysine
Figure imgf000020_0002
Reagents: a) 1) MeOH/ H+ (99.4%); 2) NaOMe, NH4CI, pH 11.5 (85%); b) Aldehyde, NaBH(OAc)3, DCE; c) arylsulfonyl chloride or substituted-arylsulfonyl chloride; TEA, CH2CI2, 3 h; d) 6N HCI, 12 h
Scheme 2 illustrates the preparation of HIV protease inhibitors bearing either a carboxylic function, compound 6, or an alcohol function, compound 8, on the final product. In other words, this scheme shows the synthesis of a L-lysine derivative or a (25) 2,6-diaminohexanol derivative
Note: a) For scheme 2, R, and R2 are as defined above b) R3 represents H, a straight or branched alkyl group of 1 to 6 carbon atoms, a phenyl or a benzyl group c) R4 is as defined above d) R5 represents an amino acid side chain as defined above. Following the indications summarized in Scheme 2, derivative 5 is linked with a substituted amino acid using NN-carbonyldiimidazole as the activating reagent to yield derivative 6 in good to excellent yields. The various N-acylated (or N-sulfonated) amino acids needed for the coupling reaction were prepared from the appropriate amino acid and acyl chloride (or sulfonyl chloride) using the Schotten-Baumen procedure. Alternatively, derivative 5 is treated with trimethylsilyl chloride in methanol (HCl generated in situ) and the resulting methyl ester intermediate is reduced with lithium aluminum hydride (LAH) in THF to afford 7 in good yields. The (2S) 2,6-diaminohexanol derivative 7 is linked to a substituted amino acid derivative as it is described above for the synthesis of derivative 6.
Scheme
Figure imgf000021_0001
LG = Leaving group
Figure imgf000021_0002
Figure imgf000021_0003
The L-lysine divative 6 can be further transformed into a variety of esters 9 as well as amide derivative 10 as shown in scheme 3. These transformations are done under standard reaction conditions. For example, the synthesis of ester 9 can be achieved upon activation of the acid 6 with DCC in the presence of a catalytic amount of NN-dimethylaminopyridine and an alcohol. The amide 10 can be obtained as described earlier for the preparation of compound 6, see scheme 2.
Note: a) For scheme 3, R]5 R2, R3, R4 and R5 are as described above b) R10 and Rn, same or different, represent an H or a straight or branched alkyl group of 1 to 4 carbon atoms.
Scheme 3
Figure imgf000022_0001
10
Scheme 4 presents a second approach for the preparation of HIV protease inhibitors of formula 6 and 8. It proceeds by using commercially available Ne-benzyloxycarbonyl-L-lysine methyl ester hydrochloride (11) as the starting material. Reductive alkylation of derivative 11 with an appropriate aldehyde and sodium cyanoborohydride provided the derivative 12. Then, sulfonation with benzenesulfonyl chloride (or substituted-benzenesulfonyl chloride) in the presence of triethylamine (or diisopropylethylamine) in dichloromethane gave compound 13 in excellent yields for the two first steps. Removal of the benzyloxycarbonyl group (Z group) by hydrogen gas in presence of 10% Pd/C yielded the free Ne-amino derivative 14 quantitatively. Acylation of 14 with a substituted amino acid N-hydroxysuccinimide ester provided derivative 15 in excellent yields. The desired HIV protease inhibitors 6 and 8 are easily obtained from the methyl ester 15 by hydrolysis with sodium hydroxide in a mixture of THF and methanol giving the acid 6 or by reduction with LAH giving the alcohol 8, both in excellent yields. It is noteworthy that, under basic hydrolysis of 15 to produce compound 6, some racemization may occur. However, it is not the case when compound 15 is reduced with LAH to give derivative 8.
Note: a) For scheme 4, R,, R2, R3, R4 and R5 are as described above
Scheme 4
MHCbz
Figure imgf000024_0001
R-i = Isobutyl
= Cyclopentylmethyl
R2CI, CH CI TEA
Figure imgf000024_0002
14 13
Figure imgf000024_0003
Scheme 5 illustrates the preparation of an anti-protease derivative using a solid phase methodology in accordance with the present invention (see example 21). Any suitable solid phase substrate could be used in such preparation (K. Burgess, Solid phase organic synthesis, Wiley-Interscience, 2000).
Note: a) For scheme 4, R, is wø-butyl, R2 is 4-methylbenzenesulfonyl, R3, R4 and R5 are as described above
This process allows the introduction of pharmacophores to a N ,N -disubstituted-L-lysine derivative (such as 16) via the N-terminal function. Thus, in scheme 5, N -isobutyl-N -(4- methylbenzenesulfonyl)-Ne-(9-fluorenylmethoxycarbonyl)-L-lysine 16 is immobilized on &p- benzyloxybenzylalcohol resin (Wang resin) in DMF for a period of 16 h. The resulting component 17 contained 0.28 meq. of L-lysine derivative / g of resin. At this stage, after removal of the Fmoc protective group under standard reaction conditions (30% piperidine in DMF see T. W. Greene and P. G. M. Wuts, Protective groups in Organic Synthesis, 3rd Edition, John Wiley & Sons, Inc. 2000), the resin can be coupled with a variety of N-acylated (or N-sulfonated) amino acids to give component 18. The N-acylated (or N-sulfonated) amino acids are activated with N-hydroxysuccinimide and DCC in DMF. Cleavage of the resin with TFA in CH2C12 leads to the desired L-lysine derivative 19.
Scheme 5
Figure imgf000026_0001
19 18
f J - Any suitable solid phase support could be used, such as, for example, polystyrene (Ps see K. Burgess, Solid phase organic synthesis, Wiley-lnterscience, 2000.
LG = Leaving group Scheme 6 illustrates the preparation of substituted glycine derivatives used for the synthesis of several HIV protease inhibitors in accordance with the present invention (see examples 114 and 158 below for specific descriptions of the synthesis of such glycine derivatives):
In scheme 6 a), N-phenylglycine 20 is treated with an excess butyllithium to give the dianion intermediate to which an appropriate alkyl halide (or arylalkyl halide or tosylate) is added and reacted for a period of 16 h. The final products 21 are obtained in good to excellent yields. An appropriate alkyl halide is defined as bearing a R3 component which can sustain strong basic reaction conditions.
In scheme 6 b), methyl bromoacetate 22 is treated with benzylamine in CH2C12 at room temperature for 16 h. The N-benzylglycine methyl ester derivative 23 was obtained in 86% yield. This intermediate can be either acylated with a carboxylic acid derivative and DCC in THF or sulfonatcd with an appropriate sulfonyl chloride and triethylamine in CH2C12 to give derivative 24 or 25 as desired in good to excellent yields.
Scheme 6
Figure imgf000028_0001
20, /V-Phenyig!ycine 21
LG = Leaving group (Ci, Br.l, OTs)
Figure imgf000028_0002
22 23
1.
Figure imgf000028_0003
25
R4 = Sulfonyl derivative Scheme 7 shows another methodology for the formation of Ne-substituted-glycyl-L-lysine HIV protease inhibitors via theNe-iodoacetyl-L-lysine derivative 26 (see example 105 for the detailed description of the synthesis of derivative 26 and its use). Thus, Nα,N -disubstituted-L-lysine derivative 5 potassium sail is initially treated with chloroacctyl chloride in the presence of DIEA in THF to give the N ,Nα-disubstituted-Ne-chloroacetyl-L-Iysine intermediate. This intermediate is transformed into the iodoacetyl derivative 26 upon treatment with sodium iodide in dr>' acetone. Compound 26 is then heated at reflux with a primary (or secondary) amine in the presence of of DIEA in THF to yield the desired Ne-substituted-glycyl-L-lysine derivative 6. In scheme 7. a primary amine is used so R3 = H and R5 = H.
Scheme 7
Figure imgf000030_0001
2. Nal, Acetone
Figure imgf000030_0002
DIEA, THF
Figure imgf000030_0003
26
R4-NH2) DIEA, THF
Figure imgf000030_0004
6
R3 and R5 = H As it can be appreciated by the skilled artisan, the above synthetic schemes are not intended to be a comprehensive list of all means by which the compounds described and claimed in this application may be synthesized. Further methods will be evident to those of ordinary skill in the art.
The compounds of this invention may be modified by appending appropriate functionalities to enhance selective biological properties. Such modifications are known in the art and include those which increase biological penetration into a given biological system (e.g., blood, lymphatic system, central nervous system), increase oral availability, increase solubility to allow administration by injection, alter metabolism and alter rate of excretion.
As discussed above, the novel compounds of the present invention are excellent ligands for aspartyl proteases, particularly HIV-1 protease. Accordingly, these compounds are capable of targeting and inliibiting late stage events in the replication, i.e. the processing of the viral polyproteins by HIV encoded protease. Compounds according to this invention advantageously inhibit the ability of the HIV-1 virus to infect immortalized human T cells over a period of days, as determined by an assay measuring the amount of extracellular p24 antigen ~ a specific marker of viral replication (see, Meek et al., Nature, 343, pp. 90-92 (1990)).
In addition to their use in the prophylaxis or treatment of HIV or HTLV infection, the compounds according to this invention may also be used as inhibitory or interruptive agents for other viruses which depend on aspartyl proteases, similar to HIV or HTLV aspartyl proteases, for obligatory events in their life cycle. Such compounds inhibit the proteolytic processing of viral polyprotein precursors by inliibiting aspartyl protease. Because aspartyl protease is essential for the production of mature virions, inhibition of that processing effectively blocks the spread of virus by inhibiting the production and reproduction of infectious virions, particularly from acutely and chronically infected cells. The compounds of this invention advantageously inhibit aspartyl proteases, thus blocking the ability of aspartyl proteases to catalyse the hydrolysis of peptide bonds.
The compounds of this invention may be employed in a conventional manner for the treatment or prevention of HIV, HTLV, and other viruses, which depend on aspartyl proteases for obligatory events in their life cycle. Such methods of treatment, their dosage levels and requirements may be selected by those of ordinary skill in the art from available methods and techniques. For example, a compound of this invention may be combined with a pharmaceutically acceptable adjuvant for administration to a virally infected patient in a pharmaceutically acceptable manner and in an amount effective to lessen the severity of the viral infection.
Alternatively, the compounds of this invention may be used in vaccines and methods for protecting individuals against viral infection over an extended period of time. The compounds may be employed in such vaccines either alone or together with other compounds of this invention in a manner consistent with the conventional utilization of protease inhibitors in vaccines. For example, a compound of this invention may be combined with pharmaceutically acceptable adjuvants conventionally employed in vaccines and administered in prophylactically effective amounts to protect individuals over an extended period of time against viral infections, such as HIV infection. As such, the novel protease inhibitors of this invention can be administered as agents for treating or preventing viral infections, including HIV infection, in a mammal.
The compounds of this invention may be administered to a healthy or HlV-infected patient either as a single agent or in combination with other antiviral agents which interfere with the replication cycle of HIV. By administering the compounds of this invention with other antiviral agents which target different events in the viral life cycle, the therapeutic effect of these compounds is potentiated. For instance, the co-administered antiviral agent can be one which targets early events in the viral life cycle, such as attachment to the cell receptor and cell entry, reverse transcription and viral DNA integration into cellular DNA. Antiviral agents targeting such early life cycle events include among others polysulfated polysaccharides, sT4 (soluble CD4) and other compounds which block binding of virus to CD4 receptors on CD4 bearing T-lymphocytes and other CD4(+) cells, or inhibit fusion of the viral envelope with the cytoplasmic membrane, and didanosine (ddl), zalcitabine (ddC), stavudine (d4T), zidovudine (AZT) and lamivudine (3TC) which inhibit reverse transcription. Other anti-retroviral and antiviral drugs may also be co- administered with the compounds of this invention to provide therapeutic treatment for substantially reducing or eliminating viral infectivity and the symptoms associated therewith. Examples of other antiviral agents include ganciclovir, dideoxycytidine, trisodium phosphonoformate, eflornithine, ribavirin, acyclovir, alpha interferon and trimenotrexate. Additionally, other types of drugs may be used to potentiate the effect of the compounds of this invention, such as viral uncoating inhibitors, inhibitors of Tat or Rev trans-activating proteins, antisense molecules or inhibitors of the viral integrase. These compounds may also be co- administered with other inhibitors of HIV aspartyl protease.
Combination therapies according to this invention exert a synergistic effect in inhibiting HIV replication because each component agent of the combination acts on a different site of HIV replication. The use of such combinations also advantageously reduces the dosage of a given conventional anti-retro viral agent that would be required for a desired therapeutic or prophylactic effect as compared to when that agent is administered as a monotherapy . These combinations may reduce or eliminate the side effects of conventional single anti-retroviral agent therapies while not interfering with the anti-retroviral activity of those agents. These combinations reduce the potential of resistance to single agent therapies, while minimizing any associated toxicity. These combinations may also increase the efficacy of the conventional agent without increasing the associated toxicity. Preferred combination therapies include the administration of a compound of this invention with AZT, 3TC, ddl, ddC, d4T or other reverse transcriptase inhibitors.
Alternatively, the compounds of this invention may also be co-administered with other HIV protease inhibitors such as Ro 31-8959 (Saquinavir; Roche), L-735,524(Indinavir; Merck), AG- 1343 (Nelfinavir; Agouron), A-84538 (Ritonavir; Abbott), ABT-378/r (Lopinavir; Abbott), and VX-478 (Amprenavir; Glaxo) to increase the effect of therapy or prophylaxis against various viral mutants or members of other HIV quasi species.
We prefer administering the compounds of this invention as single agents or in combination with retroviral reverse transcriptase inhibitors, or other HIV aspartyl protease inhibitors. We believe that the co-administration of the compounds of this invention with retroviral reverse transcriptase inhibitors or HIV aspartyl protease inhibitors may exert a substantial synergistic effect, thereby preventing, substantially reducing, or completely eliminating viral infectivity and its associated symptoms. The compounds of this invention can also be administered in combination with immunomodulators (e.g., bropirimine, anti-human alpha interferon antibody, IL-2, GM-CSF, methionine enkephalin, interferon alpha, diethyldithiocarbamate sodium, tumor necrosis factor, naltrexone and rEPO) antibiotics (e.g., pentamidine isethionate) or vaccines to prevent or combat infection and disease'associated with HIV infection, such as AIDS and ARC.
When the compounds of this invention are administered in combination therapies with other agents, they may be administered sequentially or concurrently to the patient. Alternatively, pharmaceutical or prophylactic compositions according to this invention may be comprised of a combination of an aspartyl protease inhibitor of this invention and another therapeutic or prophylactic agent.
Although this invention focuses on the use of the compounds disclosed herein for preventing and treating HIV infection, the compounds of this invention can also be used as inhibitory agents for other viruses that depend on similar aspartyl proteases for obligatory events in their life cycle. These viruses include, but are not limited to, retroviruses causing AIDS-like diseases such as simian immunodeficiency viruses, HIV-2, HTLV-I and HTLV-II. In addition, the compounds of this invention may also be used to inhibit other aspartyl proteases and, in particular, other human aspartyl proteases including renin and aspartyl proteases that process endothelin precursors.
Pharmaceutical compositions of this invention comprise any of the compounds of the present invention, and pharmaceutically acceptable salts thereof, with any pharmaceutically acceptable carrier, adjuvant or vehicle. Pharmaceutically acceptable carriers, adjuvants and vehicles that may be used in the pharmaceutical compositions of this invention include, but are not limited to ion exchangers, alumina, aluminum stearate, lecithin, serum proteins, such as human serum albumin, buffer substances such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes, such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, cellulose-based substances, poiyethyleneglycol, sodium carboxymcthylccllulosc, polyacrylates, waxes, polycthylcne- polyoxypropylene-block polymers, polyethylene glycol and wool fat. The pharmaceutical compositions of this invention may be administered orally, parenterally by inhalation spray, topically, rectally, nasally, buccally, vaginally or via an implanted reservoir. We prefer oral administration or administration by injection. The pharmaceutical compositions of this invention may contain any conventional non-toxic pharmaceutically acceptable carriers, adjuvants or vehicles. The term "parenteral" as used herein includes subcutaneous, intracutaneous, intravenous, intramuscular, intra-articular, intrasynovial, intrasternal, intrathecal, intralesional and intracranial injection or infusion techniques.
The pharmaceutical compositions may be in the form of a sterile injectable preparation, for example, as a sterile injectable aqueous or oleaginous suspension. This suspension may be formulated according to techniques known in the art using suitable dispersing or wetting agents (such as, for example, Tween 80) and suspending agents. The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent, for example, as a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that may be employed are amino acid, water, Ringer's solution and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose, any bland fixed oil may be employed including synthetic mono- or diglycerides. Fatty acids, such as oleic acid and its glyceride derivatives are useful in the preparation of injectables, as are natural pharmaceutically-acceptable oils, such as olive oil or castor oil. especially in their polyoxyethylated versions. These oil solutions or suspensions may also contain a long-chain alcohol diluent or dispersant, such as Ph. Helv. or a similar alcohol.
The pharmaceutical compositions of this invention may be orally administered in any orally acceptable dosage form including, but not limited to, capsules, tablets, and aqueous suspension and solutions. In the case of tablets for oral use, carriers that are commonly used include lactose and com starch. Lubricating agents, such as magnesium stearate, are also typically added. For oral administration in a capsule form, useful diluents include lactose and dried corn starch. When aqueous suspensions are administered orally, the active ingredient is combined with emulsifying and suspending agents. If desired, certain sweetening and/or flavoring and/or coloring agents may be added. The pharmaceutical compositions of this invention may also be administered in the form of suppositories for rectal administration. These compositions can be prepared by mixing a compound of this invention with a suitable non-irritating excipient which is solid at room temperature but liquid at the rectal temperature and therefore will melt in the rectum to release the active components. Such materials include, but are not limited to, cocoa butter, beeswax, and polyethylene glycols.
Topical administration of the pharmaceutical compositions of this invention is especially useful when the desired treatment involves areas or organs readily accessible by topical application. For application topically to the skin, the pharmaceutical composition should be formulated with a suitable ointment containing the active components suspended or dissolved in a carrier. Carriers for topical administration of the compounds of this invention include, but are not limited to, mineral oil, liquid petroleum, white petroleum, propylene glycol, polyoxyethylene or polyoxypropylene compound, emulsifying wax and water. Alternatively, the pharmaceutical compositions can be formulated with a suitable lotion or cream containing the active compound suspended or dissolved in a carrier. Suitable carriers include, but are not limited to, mineral oil, sorbitan monostearate, polysorbate 60, cety 1 esters wax cetearyl alcohol, 2-octyldodecanol, benzyl alcohol and water. The pharmaceutical compositions of this invention may also be topically applied to the lower intestinal tracl by recta] suppository formulation or in a suitable neat formulation. Topically-transdermal patches are also included in this invention.
The pharmaceutical compositions of this invention may be administered by nasal aerosol or inhalation. Such compositions are prepared according to teclmiques well-known in the art of pharmaceutical formulation and may be prepared as solutions in saline employing benzyl alcohol or other suitable preservatives, absorption promoters to enhance bioavailability, fluorocarbons, and/or other solubilizing or dispersing agents known in the art.
Dosage levels of between about 0.01 and about 25 mg/kg body weight per day, preferably between about 0.5 and about 25 mg/kg body weight per day of the active ingredient compound are useful in the prevention and treatment of viral infection, including HIV infection. Typically, the pharmaceutical compositions of this invention will be administered from about 1 to about 5 times per day or alternatively, as a continuous infusion. Such administration can be used as a chronic or acute therapy. The amount of active ingredient that may be combined with the carrier materials to produce a single dosage form will vary depending upon the patient treated and the particular mode of administration. A typical preparation will contain from about 5%to about 95% active compound (w/w). Preferably, such preparations contain from about 20% to about 80% active compound.
Upon improvement of a patient's condition, a maintenance dose of a compound, composition or combination of this invention may be administered if necessary. Subsequently, the dosage or frequency of administration, or both, may be reduced, as a function of the symptoms, to a level at which the improved condition is retained. When the symptoms have been alleviated to the desired level, treatment should cease. Patients may, however, require intermittent treatment on a long-term basis, upon any recurrence of disease symptoms.
As the skilled artisan will appreciate, lower or higher doses than those recited above may be required. Specific dosage and treatment regimen for any particular patient will depend upon a variety of factors, including the activity of the specific compound employed, the age, body weight, general health status, sex, diet, time of administration, rate of excretion, drug combinati on. the severity and course of the infection, the pal ient' s disposition to the infection and the judgment of the treating physician.
The compounds of this invention are also useful as commercial reagents which effectively bind to aspartyl proteases, particularly HIV aspartyl protease. As commercial reagents, the compounds of this invention, and their derivatives, may be used to block proteolysis of a target peptide by an aspartyl protease, or may be derivatized to bind to a stable resin as a tethered substrate for affinity chromatography applications. These and other uses which characterize commercial aspartyl protease inhibitors will be evident to those of ordinary skill in the art. Enzymatic assay for determining the inhibition constant (Ki) of synthetic compounds targeting the HIV protease
This is a fluorometric assay based on the cleavage by protease of a substrate carrying a donor group (EDANS) and an acceptor group (DABCYL) on each side of the cleavage site, interacting together through fluorescence resonance energy transfer (FRET) as described by Matayoshi et al. (Science 247:954-954, 1990).
After calculation of Vo and Vi, the inhibition constant (Ki) of the compound is determined using the equation of Henderson.:
Vo [I] Where Kiapp = 1 + • Ki = i Kiapp 1 + [S]
Km
where Vo = the enzyme's initial velocity
Vi = the enzyme velocity' in the presence of the inhibitory compound, [ I ] = inhibitor concentration, [ S ] = substrate concentration, Km - Michaelis-Menten constant and Kiapp = apparent Ki
Graphs are traced and the Ki determined using GraphPad Prism software v. 3.0.
The compounds listed in Tables 1 and 2 were prepared by following Scheme 1, 2, 3, 4, 5, 6 or 7; the numbers of the compounds listed in the table correspond to the example numbers presented in the experimental section (see examples below). The activities of the compounds are also listed in the same tables demonstrating their potential usefulness. In Table 1 are shown compounds of formula I wherem Y, n, Cx, Ri5 R2, R3, Rj and R5 are as presented in Table 1. In Table 2 are shown compounds of formula II wherem Y, n, Cx, R„ R2, R4 and ^ are as presented in Table 2.
31 escription herein, the following abbreviations are used:
Abbreviation Meaning
Ac Acetyl
AcOH Acetic acid
ARC AIDS-related complex
AIDS Acquired Immunodeficiency Syndrome
AZT 3 - Azido-3 -deoxythymine (Zidovudine)
Bn benzyl
Boc tert-Butoxycarbonyl i-Bu iso-Butyl t-Bu tert-Butyl
CAM Cerium ammonium molybdate
DABCYL 4-[[4'-(dimethylamino)phenyl]azo]benzoic acid
DCC Dicyclohexylcarbodiimide
DCE Dichloroethane
DCM Dichloromethane
DMAP NN-dimethylammopyridine
DIEA NNDiisopropylethylamine
DMF Dimethylform amide
DNA Deoxyribonucleic acid
EDANS 5-[(2'-aminόethyl)amino]naphthalene sulfonic acid
EDC 1 -Ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride
EtOAc Ethyl acetate
EtOH Ethyl alcohol
Fmoc 9-Fluorenylmethoxy carbonyl g Gram
HIV-1 , -2 Human immunodeficiency virus type 1 , type 2 HOBt 1 -Hydroxybenzotriazole HPLC High performance liquid chromatography HTLV-I, -II Human T-cell lymphotropic virus type I, type II IL-2 lnterleukin-2
Kg Kilogram
LAH Lithium aluminum hydride
LC-MS Liquid chromatography-mass spectrometry
M Molar
MeOH Methyl alcohol mg Milligram
MP Melting point min Minute mol Mole mL Milliliter mmol Millimole nM Nanomolar z'-Pr wo-Propyl rEPO Recombinant erythropoietin
RNA Ribonucleic acid
3TC 2',3'-Dideoxy-3-thiacytidine
TFA Trifluoroacetic acid
H- TFA Trifluoroacetic acid ammonium salt
THF Tetrahydrofuran
Z Benzyloxycarbonyl
EXAMPLES
In order that this invention be more fully understood, the following examples are set forth. These examples are for the purpose of illustration only and are not to be construed as limiting the scope of the invention in any way. Materials and Methods
Analytical thin layer cliromatography (TLC) was carried out with 0.25 mm silica gel E. Merck 60 F254 plates and eluted with the indicated solvent systems. Preparative chromatography was perfomied by flash chromatography, using silica gel 60 (EM Science) with the indicated solvent systems and positive air pressure to allow proper rate of elution. Detection of the compounds was carried out by exposing eluted plates (analytical or preparative) to iodine, UV light and/or treating analytical plates with a 2% solution ofp-anisaldehyde in ethanol containing 3% sulfuric acid and 1% acetic acid followed by heating. Alternatively, analytical plates can be treated with a 0.3% ninhydrin solution in ethanol containing 3% acetic acid and/or a CAM solution made of 20 g (NH4)6Mo7O24 and 8.3 g Ce(S04)2 polyhydrate in water (750 mL) containing concentrated sulfuric acid (90 mL).
Preparative HPLC were perform on a Gilson apparatus equipped with a Cl 8 column, a215 liquid handler module and 15 mL/min capacity head pumps. The HPLC is operated with a Gilson UniPoint System Software. A solvent gradient was used starting from H2O/CH3CN (95%:5%) to 100% CH3CN over 25 min, and 100% CH3CN for a further 20 min to clean the column.
Unless otherwise indicated, all starting materials were purchased from a commercial source such as Aldrich Co. or Sigma Co.
Nuclear magnetic resonance (NMR) spectra were recorded on a Bruker AMX 500 equipped with a reversed or QNP probe. Samples were dissolved in deuterochloroform (CDC13), deuteroacetone (acetone-d6) or deuterodimethylsulfoxide (DMSO-d6) for data acquisition using tetramethylsilane as internal standard. Chemical shifts (δ) are expressed in parts per million (pp ), the coupling constants (J) are expressed in hertz (Hz) whereas multiplicities are denoted as s for singlet, d for doublet, dd for doublet of doublets, t for triplet, q for quartet, m for multiplex, and br s for broad singlet. GENERAL PROCEDURES
A. Preparation of Ar-acylated (or N-sulfonated) amino acids (Schotten-Baumen procedure)
To a solution of an amino acid (10 mmol) in 25 mL IN ΝaOH and 5 mL saturated Νa2CO3 (resulting solution at pH 10) was added an acyl chloride (or a sulfonyl chloride, or a chloroformate) (12 mmol) dissolved in 10 mL acetone over a period of 20 min. Afterwards, the reaction mixture was stirred at room temperature for 2 h. The alkaline solution was extracted once with ether (50 mL) and the aqueous phase was acidified with IN HCl to form a pasty oil. This was extracted twice with 20 mL CHC13, and the combined organic phases were washed with 10 mL IN HCl. The organic phase was dried over MgSO4_ filtered and evaporated to an oil which crystallized on standing. The solid was recrystallized from either dichloromethane, ether, hexanes or without solvent as indicated in each specific example. The purity was evaluated by LC-MS and/or Η NMR and was found to be ranging from 85 to 99%.
B. Coupling reaction of N-acylated (or N-sulfonated) amino acid with the Ne-ΝH2 of a L- lysine derivative
Depending on the nature of the reagents, various methods were used to link the two amino acid portions together.
a) NN-carbonyldiimidazole method
Nα-isobutyl-Λ7α-(4-methylbenzenesulfonyl)-L-lysinehydrochloride (or other L-lysine derivative) (100 mg, 0.25 mmol) was weighed in the Bohdahn robotic reaction vessels. The solid was then added to 1 L 3.3M Cs2CO3 solution to which 2 mL of THF was added. The tube was then stirred vigorously. The resulting mixture was treated with N-acylated (or N-sulfonated) amino acid (0.3 mmol) activated by NN-carbonyldiimidazole (0.3 mmol) dissolved in THF (1 mL). The stirring was continued for 2 h. Afterwards, EtOAc (3 mL) was added and the organic phase was removed. The organic phase was washed with IN HCl and again separated. Evacuation of the solvent gave a crude product which was resolved by HPLC. The yield of the reactions will be indicated in each specific example.
b) Solid phase method
Preparation of solid-phase bound N -isobutyl-N -(4-methylbenzenesulfonyl)-Ne-(9- fluorenylmethoxycarbonyP-L-lysine
Λr -isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-(9-fluorenylmethoxycarbonyl)-L-lysine (1.51 g, 2.6 mmol) was dissolved in DCM (70 mL) containing DCC (1.5 g). The solution was stirred at room temperature for 8 h and then filtered. The filtrate was added to 5.0 g dried washed Wang resin (0.73 meq/g) to which 150 mg NN-dimethylaminopyridine (DMAP) was added. The suspension was stirred at room temperature for 12 h. Then, the resin was filtered and washed successively with DCM (100 mL, 2X), 1 :1 DCM : MeOH (100 mL, 3X), MeOH (50 mL, 2X) and ether (100 mL). The resin was again swollen in DCM to which acetic anhydride (20 mL) was added. It was left to stand for 3 h and then filtered and washed as above. The resulting resin was dried at room temperature in a dessicator in vac o. The resulting resin (5.92 g) contained 0.28 mcq/g L-lysine derivative.
ΝB: Same preparation for solid-phase bound Nα-isobutyl-N -(4-nifrobenzenesulfonyl)-Ne- (9-fluorenylmethoxycarbonyl)-L-lysine (see example 28).
Deprotection
In a typical experiment, 450 mg (0.125 mmol) of resin was added to a syringe type reaction vessel with Teflon frits and stopcock. The resin was swollen with DCM and washed after 15 min. It was treated with 30% piperidine in DMF (4 mL) and left for 15 min before being successively washed with DMF (5 mL, 2X), DCM (5 mL, 4X), and ether (5 mL,.4X). This process was repeated once.
Coupling
In a typical experiment, 0.5 mmol of N-acylated (or N-sulfonated) amino acid was added to a solution of N-hydroxysuccinimide (0.5 mmol) and DCC (0.5 mmol) in DMF (3 mL). The acid was activated for 3 h and filtered directly into the resin containing vessel. The coupling reaction was allowed to proceed for 12 h at room temperature. The resin was then washed successively with DCM, MeOH and ether as described above then dried in vαcuo.
Cleavage
The dried resin was swollen with DCM, filtered and treated with 95% TFA (4 mL). The resulting mixture was stirred, for a period of 3 h. Then, the solution was filtered off and evaporated. The residue was triturated with ether and the pasty solid placed under high vacuum for 4 h. The solid was purified by preparative HPLC to give the final coupled product. The yield of the reactions will be indicated in each specific example.
c) Dicyclohexylcarbodiimide (DCC) method
Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L -lysine potassium salt (or other L-lysine potassium salt derivative) (1 mmol) was weighed in a round bottom flask and was suspended in THF (20 L). This suspension was treated with a IN ΝaOH (1.5 mL) to pH 10. In another reaction vessel, a solution of N~protected amino acid (1 mmol) in THF (25 mL), was treated with 115 mg of N-h droxysuccinimide (1 mmol) and 206 mg DCC (1 mmol). The resulting mixture was stirred at room temperature for 4 h. Afterwards, the precipitate was filtered off and the filtrate was added to the initial suspension with vigorous stirring. After 2 h, a 2 mL aliquot of water was added resulting in a clear solution. The stirring was continued for 12 h. Then, EtOAc (50 mL) was added and the organic phase was washed successively with lΝΝaOH (50 mL), with IN HCl (50 mL) and finally with brine (50 mL). The organic phase was removed and dried withΝa2S04. The solvent was evaporated and the product was purified (in whole or in part) by preparative HPLC or by trituration with ether. The yield of the reactions will be indicated in each specific example.
d) l-Ethyl-3-(3-dimethyIaminopropyI)carbodiimide hydrochloride (EDC) method
A solution of N -isobutyl-Nc -(4-methylbenzenesulfonyl)-L-lysinol (1.63 g) in EtOAc (16 mL) was prepared (100 g/ mL, 0.29 mmol/mL). A second solution containing EDC (2.5 g) and HOBt (1.34 g) in DMF was prepared (0.5 mmol / mL). To Bohdan robotic test tubes were then added a series of N-substituted amino acids (0.5 mmol) to which a 1 mL aliquot of the EDC : HOBt solution was added. After 20 min, a 1 mL aliquot of the lysinol solution was added. The resulting solutions were stirred at room temperature for 6 h. Afterwards, a 5 mL aliquot of 10% citric acid aqueous solution was added to each test tube and the solutions were extracted with EtOAc (50 mL). The organic phase were evaporated and the residue obtained in each test tube was purified by HPLC. The yield of the reactions will be indicated in each specific example.
C. Removal of the Λ-ter/-butoxycarbonyl (Boc) group
To a series of Boc protected products (100 mg) in Bohdan test tubes was added 2 mL of CH2C12 : TFA (1 :1). Gas evolution was observed and the solutions were stirred for 20 min. The solvents were evacuated and the resulting thick oil was triturated with cold ether. The ether was decanted away and the remaining products were placed in a high vacuum desiccator for 12 h to give solid foams. The yield of the reactions will be indicated in each specific example.
D. Sodium salt formation
The L-lysine product (100 mg) was dissolved in MeCΝ (1 mL). The solution was added to 5 mL H20 to form a turbid suspension. A IN ΝaOH solution (1 mol eq) was slowly added to' the turbid suspension which became clear. The solution was frozen solid and lyophilized to yield a white powder (100%).
E. Catalytic hydrogenation
To a series of nitro compounds (100 mg) dissolved in argon saturated MeOH (10 mL) was added 10% Pd/C (50 mg) followed by formic acid 98% (0.1 mL). The suspensions were saturated with H2 and kept under positive pressure using a H2 filled balloons. After 4-6 h stirring, the H2 was purged out and the solutions were filtered through thin pads of celite. The clear solutions were then evacuated, triturated with ether and resolved by preparative HPLC. The yield of the reactions will be indicated in each specific example. F. General procedure for the thiomidation reaction
To a stirred solution of amide (lmmol) in dry THF (10 mL) was added Lawesson's reagent (606 mg, 1.5 mmol). The reaction was stirred overnight then concentrated and purified by flash chromatography using hexane/EtOAc as eluent to afford the desired thioamide.
G. General Procedure for the reduction of esters with LiAlH4
To a stirred solution of the ester (1 mmol) in THF was added at 0°C LiAlH4 (1.5 mmol). The mixture was stirred at room temperature for 3 h. The hydride excess was neutralised with HCl IN and the reaction was extracted with EtOAc. The organic phase was dried (MgSO4) and concentrated. The crude was purified by flash cliromatography.
H. Substitution reaction on an iodoacetamide derivative
To a solution of Nα-isobutyl-N -(4-methylbenzenesulfonyl)-Ne-iodoacetyl-L-lysine (1.0 eq.) in THF (10. mL) was added DIEA (2.0-3.0 eq.) and an amine derivative (2.0-4.0 eq.). The reaction mixture was stirred overnight at room temperature. Then, a 2Ν HCl solution (2 mL) was added and the resulting mixture was extracted with EtOAc (20 L, 3X). The organic phase was dried MgSO4 and evaporated to an oil. The crude material was purified by preparative HPLC.
Specific examples for the preparation of derivatives of general formula I
The following compounds were prepared either from a L-amino acid or, when indicated, from a derivative of a D-amino acid using the procedures summarized in "schemes 1, 2, 3, 4, 5, 6 and 7. Example 1. Preparation of N -isoburyl-Nα^-methylbenzenesulfony^-Ne-fN'α^- m ethylb enzen esulf ony l)-L-phenylalanyl] -L-ly sine
Step A. Preparation of L-lysine methyl ester dihydrochloride • MeOH (J. Org. Chem. 44, 4841 (1979))
To a stirred suspension of L-lysine monohydrochloride (190.7 g, 1.08 mol) in MeOH (3 L) was added (via a cannula) trimethylsilylchloride (350 mL). The mixture quickly became clear and homogeneous. The solution was stirred at reflux for 3 h and then at room temperature for 2 h. The reaction flask was left overnight in a refrigerator cooled to - 75 °C. The large crystals obtained were filtered, washed with cold MeOH (100 mL) and dried in vacuo for 24 h at room temperature. L-lysine methyl ester dihydrochloride • MeOH (275.8 g) was obtained in 99.4% yield.
!HΝMR (DMSO-dό): δ 1.36 (m, 1H), 1.45 (m, 1H), 1.58 (m, 2H), 1.81 (m, 2H), 2.74 (br s, 2H), 3.11 (s, 3H), 3.72 (s, 3H), 3.94 (t, J= 4.0, 1H), 8.12 (br s, 3H), 8.72 (br s, 3H).
StepB. Preparation of L-α-amino-e-caprolactam hydrochloride (J. Org. Chem. 44, 4841 (1979))
Sodium methylate 58.73 g (1 mole) was dissolved in cold MeOH (1 L). About one half of this solution was cannulated into a solution of L-lysine methyl ester dihydrochloride • MeOH (132.5 g, 0.5 mole) in 1 L MeOH. The suspension was allowed to warm and dissolved. The remainder sodium methylate was added with concurrent apparition of NaCl. The mixture was then allowed to reflux for 4 h, after which 5 g of NH4C1 was added. The solution then sat at RT for 18 h and was filtered through celite. Evaporation of the MeOH resulted in a thick opaque syrup. The excess NaCl was removed by redissolving the mixture in boiling glyme (100 L, 2X), filtering through celite and evaporating in vacuo. The resulting clear oil was taken up in ethanol and acidified with 12 N HCl. Cooling gave a mass of fline white needles which were filtered and dried in vacuo to yield 69.71 g, 85% of the title compound. MP 301-306 °C [α]d = - 24.8 (c = 3.4, IN HCl). Η NMR (DMSO-d6): δ 1.17 (q, J = 12.6, 1H), 1.45 (q, J = 12.6, 1H), 1.58 (q, J= 12.6, 1H), 1.71 (d, J= 12.6, 1H), 1.86 (d, = 12.6, 1H), 1.94 (d, J=- 12.6, 1H), 3.03 (m, 1H), 3.15 ( , 1H), 4.03 (d, J= 12.6, 1H), 8.12 (br s, 1H), 8.22 (br s, 3H). 13C NMR (DMSO-d6): δ 28.2, 29.7, 29.9, 41.6, 53.4, 173.2. LC-MS: 129.1 (M + H)\ 99% pure.
Step C. Preparation of N -isobutyl-L- -amino-e-caprolactam
L-α-amino-e-caprolactam (60.0 g, 0.47 mol) was dissolved in dichloroethane (DCE, 100 mL) containing isobutyraldehyde (37.0 g, 0.5 mole) and stirred until the heat evolved was dissipated. Then, DCE (2 L) and AcOH (35 mL) were added to the solution followed by 0.5 mole of powdered ΝaBH(OAc)3. The slightly turbid mixture was stirred at 60 °C for 2 h, and at room temperature for 12 h. The solution was treated with IM K2CO3 (1 L) and stirred for a further 2 h. The DCE layer was dried with MgSO4, filtered and evaporated. The oil thus obtained crystallizes slowly on standing (87 g, 94.5%) and was used without further purification in the next step. MP 52-54 °C. A small sample was converted to the hydrochloride salt by adding the solid to a solution of IN HCl in 95% EtOH.
'H NMR (CDC13): δ 0.93 (d, J= 6.5, 3H), 0.97 (ά, J= 6.5, 3H), 1.39 (t, J= 9.8, 1H), 1.47 (m, 1H), 1.78-1.65 (m, 2H), 2.00-1.93 (m, 2H), 2.32-2.2 (m, 2H), 2.38 (t, = 9.7, 1H), 3.16 (m, 3H), 6.62 (s. 1H (NH)).
Step D. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesuIfonyl)-L-α-amino-e-caprolactam
The compound prepared in step C of this example (10.0 g, 51 mmol, free base) was dissolved in DCM (100 mL) and treated with diisopropylethylamine (10 mL) followed by freshly recrystallized 4-methylbenzenesulfonyl chloride (11.4 g, 57.3 mmol). The mixture was stirred overnight (TLC shows the reaction to be complete after 2 h). The solution was extracted with IN HCl and the organic layer was dried and evaporated. Then, the residue was dissolved in boiling CHC13 (5 mL), diluted with hexanes (200 mL) and placed in the refrigerator for 3 h. The precipitated product was filtered off and air dried giving 15.5 g of pure product. MP 49-51 °C 'H NMR (CDCI3): δ 0.74 (d, J= 6.2, 3H), 0.80 (d, J= 6.2, 3H), 1.12 (q, J= 8.3, 1H), 1.56-1.73 (m, 4H), 1.84-1.87(m, 1H), 1.96-1.99 (m, 1H), 2.33 (s, 3H), 2.86-2.89 ( , 1H), 2.97-2.98 (m 1H), 3.1-3.06 (m, 2H), 3.21-3.26 (m, 1H), 4.48 (d, J= 10.6, 1H), 5.7 (s, 1H (NH)), 7.29 (d, J= 7.7, 2H), 7.59 (d, J= 7J, 2H).
Step E. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine hydrochloride ,
A mixture of Λrα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L- -amino-e-caprolactam (13.5 g, 40 mmol), AcOH (4 mL) and 6Ν HCl (200 mL) was refiuxed for 12 h until all solids had disappeared. Afterwards, the solution was evaporated to give 11.0 g, 77% of the hydrochloride salt.
!H NMR (DMSO-d6): δ 0.72 (dd, J= 5.8, 6.4, 6H), 1.13-1.17 (m, 2H), 1.17-1.24 (m 2H), 1.42- 1.48 (m, 2H), 2.3 (s, 3H), 2.67 (t, = 7.2, 2H), 2.80-2.91 (m, 2H). 4.13 (t, J= 7.2, 1H). 7.22 (d, = 8.5, 2H), 7.64 (d, J= 8.5, 2H).
Step F. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-[N'α-(4- methylbenzenesulfonyl)-L-ρhenylalanyl]-L-lysine
A suspension of Λ^α-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine hydrochloride (600 mg) in THF (20 mL) was treated with a IN ΝaOH (1.5 mL) to pH 10. A solution of commercially available Nα-(4-methylbenzenesulfonyl)-L-phenylalamne acid chloride (250 mg) in dry THF (20 mL) was added to the suspension and stirred for 2 h. Afterwards, water (2 mL) was added resulting in a clear solution. The reaction mixture was stirred for 12 h. Then, EtOAc (30 L) was added and the organic phase was washed with IN HCl. The organic phase was removed. Evaporation of the solvent gave a crude product which was triturated with ether to yield 750 mg (76%) of the title compound.
Η ΝMR (CDC13): δ 0.77 (d, ./= 6.3, 3H), 0.80 (d, J= 6.3, 3H), 1.00-1.11 (m, 4H), 1.23-1.25 (m, 1H), 1 J0-1 J4 (m, 1H), 1.89-1.93 (m, 1H), 2.30 (s, 3H), 2.32 (s, 3H), 2.59-2.67 (m, 2H), 2.87 and 2.93 (ABX, J= 14.1, 4.2, 2H), 3.85 (t, J= 5.9, 1H), 3.63 (t, J= 6.9, 1H), 6.90-7.10 (m, 7H), 7.24 (d, J= 8.0, 2H), 7.44 (d, J= 8.1, 2H), 7.73 (d, J= 8.1, 2H). LC-MS: 656.2 (M - H)\ 98% pure.
Example 2. Preparation of Nα-isobutyl-N -(4-methylbenzenesulfonyl)-Ne-[N' -(4- methylbenzenesulfonyl)-L-tryptophanyI]-L-lysine
Step A. Preparation of Nα-(4-methylbenzenesulfonyl)-L-tryptophan
L-tryptophan was reacted with 4-methylbenzenesulfonyl chloride under the conditions used in general procedure A giving the title compound which was recrystallised from DCM (58%).
]H ΝMR (CDC13): δ 2.33 (s, 3H), 2.9-3.11 (m, 2H), 3.91 (t, J= 7.0, 1H), 6.86-7.01 (m, 3H), 7.25 (d, J= 6.9, 2H), 7.34 (t, J= 6.8, IE), 7.45 (d, J= 6.9, 2H), 8.15 (d, J= 6.1, 1H). LC-MS:
357 (M - H)", 99% pure.
Step B. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-[N'α-(4- methylbenzenesulfonyl)-L-ttyptophanyl]-L-lysine
The title compound was prepared fromΛ;c.-isobutyl-N -(4-methylbenzenesulfonyl)-L-lysine (150 mg, 0.42 mmol, example 1, step E) as described in general procedure Be using N -(4- methylbenzenesulfonyl)-L-tryptophan (180 mg, 0.5 mmol) which was prepared in step A of this example. The final product was triturated with ether to yield 180 mg (69%) of the desired material.
!H ΝMR (DMSO-d6): δ 0.77 (d,J= 6.3, 3H), 0.80 (d, J= 6.3, 3H), 1.00-1.11 (m, 4H), 1.32-1.35 (m, 1H), 1 J0-1 J4 (m, 1H), 1.83-1.88 (m, 1H), 2.26 (s, 3H), 2.34 (s, 3H), 2.59-2.67 (m, 2H), 2.92 and 2.87 (ABX, J = 13.1 , 2.8, 2H), 3.80 (t, = 6.5, 1H), 4.11 (t, J= 7.2, 1H), 6.85 (t, J = 7.1 , 1H), 7.00 (t, -7 = 4.0, 2H), 7.10 (d, = 7.1, 2H), 7.28 (d, J= 4.0, 1H), 7.33 (m, 3H), 7.43 (d, = 7.1, 2H), 7.60 (d,J= 7.1 , 2H), 7.71 (t,!/= 5.4, 1H), 7.80 (d, .7= 8.0, 1H). LC-MS: 695.2 (M - H)-, 99% pure. Example 3. Preparation of N -isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-[N' -(4- acetamidobenzenesulfonyl)-L-phenylalanyl]-L-lysine
Step A. Preparation of Nα-(4-acetamidobenzenesulfonyl)-L-phenylalanine
L-phenylalanine was reacted with 4-acetamidobenzenesulfonyl chloride under the conditions used in general procedure A giving the title compound which was recrystallised from DCM (52%).
LC-MS: 362 (M - H)\ 95% pure.
Step B. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-[N'α-(4- acetamidobenzenesulfonyl)-L-phenylalanyl]-L-lysine
The title compound was prepared from Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine (100 mg, 0.29 mmol. example 1, step E) as described in general procedure Be using N -(4- acetamidobenzenesulfonyl)-L-phenylalanine (102 mg, 0.29 mmol) which was prepared in step A of this example. The final product was triturated with ether to yield 101 mg (57%) of the desired material.
LC-MS: 699.2 (M - H)', 95% pure.
Example 4. Preparation of Nα-isobutyl-Nα-(4-methyIbenzenesuIfonyl)-Ne-(N'α- benzenesulfonyl-L-tryptophanyI)-L-lysine
Step A. Preparation of Nα-benzenesulfonyl-L-tryptophan
L-tryptophan was reacted with benzenesulfonyl chloride under the conditions used in general procedure A giving the title compound which was recrystallised from DCM (26%).
'H ΝMR (CDC13): δ 2.86-3.26 (m, 2H), 3.93 (t, J= 5.0, 1H), 6.92-7.00 (m, 3H), 7.28 (d, = 7.0, 2H), 7.30-7.34 (m, 3H), 7.55 (d, J= 6.0, 1H), 8.24 (d, J= 6.0, 1H). LC-MS: 343 (M - Hr, 99% pure.
Step B . Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-(N α-benzenesulfonyl-L- tryptophanyl)-L-lysine
The title compound was prepared from Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-ly sine (1.0 g, 2.9 mmol, example 1 , step E) as described in general procedure Be using Nα-benzenesulfonyl- L-tryptophan (1.72 g, 5 mmol) which was prepared in step A of this example. The final product was triturated with ether to yield 1 J g of the crude material. Purification of 500 mg of the crude material by HPLC gave 322 mg (64%) of pure adduct.
T-I ΝMR (CDC13): δ 0.78 (d, = 6.3, 3H), 0.81 (d, = 6.3, 3H), 0.94-1.03 (m, 4H), 1.32-1.35 (m, 1H), 1.46-1.49 (m, 1H), 1.83-1.88 (m, 1H), 2.35 (s, 3H), 2.82-2.99 (m, 4H), 3.95 (t, J= 6.5, 1H), 4.21 (t, J= 7.2, 1H), 6.85 (t, J= 4.5, 1H), 7.09 (t, J= 4.5, 1H), 7.23-7.31 (m, 6H), 7.42 (t, J= 4.5, 1H), 7.60 (d, J=> 6.8, 2H), 7.73 (d, J= 6.8, 2H). LC-MS: 681.2 (M - H)\ 99% pure.
Example 5. Preparation of Nα-isobutyl-Nα-(4-aminobenzenesulfonyl)-Ne-[N'α-(4- aminobenzenesulfonyi)-L-tryptophanyl]-L-rysine
Step A. Preparation of Nα-(4-nitrobenzenesulfonyl)-L-tryptophan
L-tryptophan was reacted with 4-nitrobenzenesulfonyl chloride under the conditions used in genera] procedure A giving Nα-(4-nitroberιzenesulfonyl)-L-tryptophan which was recrystallised from DCM (56%).
LC-MS: 388 (M - H)\ 99% pure.
Step B. Preparation of Nα-isobuty'l-Nα-(4-nitrobenzenesulfonyl)-L-α-amino-e-caprolactam Nα-isobutyl-L-α-amino-e-caprolactam (example 1, step C) (4.14 g, 21.1 mmol, free base) was dissolved in DCM (50 mL) and treated with diisopropylethylamine (6.0 mL, 30 mmol) followed by freshly recrystallized 4-nitrobenzenesulfonyl chloride (5.09 g, 21 J mmol). The mixture was stirred overnight (TLC shows the reaction to be complete after 2 h). The solution was extracted with IN HCl and the organic layer was dried and evaporated. Then, the residue was dissolved in boiling MeOH (250 mL) and placed in the refrigerator for 3 h. The thin needles obtained were filtered off and air dried giving 6.9 g (83%) of pure product. MP 152-154 °C
'H ΝMR (CDC13): δ 0.93 (d, J= 6.0, 3H), 0.96 (d, J= 6.0, 3H), 1.39 (t, J= 12.0, 1H), 1.65- 1.85 (m, 3H), 2.08-2.18 (m, 3H), 3.06 (dd, J= 14.2, 8.5, 1H), 3.35 (dd, J= 14.2, 8.5, 1H), 4.65 (d, J= 8.7, 1H), 5.7 (s, 1H (ΝH)), 7.92 (d, J= 8.8, 2H), 8.3 (d, = 8.8, 2H).
Step C. Preparation of Nα-isobutyl-Nα-(4-nitrobenzenesulfonyl)-L-lysine hydrochloride
A mixture of Λfα-isobutyl-Λ7'α-(4-nitrobenzenesulfonyl)-L- -amino-e-caprolactam (1.0 g, 2J mmol), AcOH (4 mL) and 6Ν HCl ( 10 mL) was ref luxed for 12 h until all solids had disappeared. Afterwards, the solution was evaporated to give 1.12 g, 100% of the hydrochloride salt.
[α]d = -16.7 (c = 0.36 in MeOH); 'HNMR (DMSO-d6): δ 0.79 (d,/= 6.8, 3H), 0.86 (d, 7= 6.8, 3H), 1.25 (t, J= 11.9, 2H), 1.32-1.28 (m 2H), 1.58-1.45 (m, 2H), 1.85-1.75 (m, 2H), 2.7 (m, 3H (NH)); 2.83 -2.87 (m, 1H), 3.03-3.07 (m, 1H), 4.21 (t, J= 10.1, 1H), 8.10 (d,J= 7.9, 2H), 8.37 (d, J= 7.9, 2H).
Step D. Preparation of Nα-isobutyl-Nα-(4-aminobenzenesulfonyl)-Ne-[N'α-(4- aminobenzenesulfonyl)-L-tryptophanyl]-L-lysine
The title compound was prepared from Nα-isobutyl-Nα-(4-nitrohenzenesulfonyl)-L-lysine hydrochloride (200 mg, 0.52 mmol, step C) as described in general procedure Be using Nα-(4- nitrobenzenesulfonyl)-L-trγptophan (300 mg, 0.8 mmol) which was prepared in step A of this example. The intermediate derivative was reduced following the indications of general procedure E. The final product was purified by HPLC to give 101 mg (53%) of pure adduct. !H NMR(DMSO-d6): δ 0.73 (d,-/= 6.3, 3H), 0.75 (d,J= 6.3, 3H), 1.00-1.11 (m,4H), 1.32-1.35 (m, 1H), 1.66-1.69 (m, 1H), 1.83-1.88 (m, 1H), 2.48 (br s, 6H), 2.59-2.67 (m, 2H), 2.84-2.96 (m, 211), 3.80 (t, J= 6.5, 1H), 4.01 (t, J = 7.2, 1H), 6.46 (d, J= 7.1, 2H), 6.52 (d, J= 7.1, 2H), 6.85 (t, J= 4.0, 1H), 7.09 (t, = 4.0, 2H), 7.28 (d, J= 7.1, 1H), 7.33 (d, J= 7.1, 2H), 7.60 (t, J= 4.0, 1H). LC-MS: 697.2 (M - H)", 98% pure.
Example 6. Preparation of Nα-isobutyl-Nα-(4-methyIbenzenesulfonyI)-Ne-[N,α-(4- nitrobenzenesulfonyl)-L-tryptophanyl]-L-lysine
The title compound was prepared from Λrα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine (100 mg, 0.29 mmol, example 1, step E) as described in general procedure Be using Nα-(4- nitrobenzenesulfonyl)-L-tryptophan (120 mg, 0.3 mmol) which was prepared in step A of example 5. The final product was purified by HPLC to give 66 mg (36%) of pure adduct.
LC-MS: 726.2 (M - H)\ 99% pure.
Example 7. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-[N'α-(4- methylbenzenesulfonyl)-D-phenyIaIanyI]-L-Iysine
Step A. Preparation of Nα-(4-methylbenzenesulfonyl)-D-phenylalanine
D-phenylalanine was reacted with 4-methylbenzenesulfonyl chloride under the conditions used in general procedure A giving the title compound which was recrystallised from ether (18%).
LC-MS: 318 (M - H)\ 98% pure.
Step B. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-[N'α-(4- methylbenzenesulfonyl)-D-phenylalanyl]-L-lysine
The title compound was prepared from Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine (100 mg, 0.29 mmol, example 1 , step E) as described in general procedure Be using N -(4- methylbenzenesulfonyl)-D-phenylalanine (160 mg, 0.5 mmol) which was prepared in step A of this example. The final product was triturated with ether to yield 49 mg (29%) of the desired material.
LC-MS: 656.2 (M - H)\ 99% pure.
Example 8. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-(N'α- benzenesulfonyl-L-phenylalanyl)-L-lysine
Step A. Preparation of Nα-benzenesulfonyl-L-phenylalanine
L-phenylalanine was reacted with benzenesulfonyl chloride under the conditions used in general procedure A giving the title compound which was recrystallised from ether (50%).
LC-MS: 349 (M - H)\ 95% pure.
Step B. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-(N'α-benzenesulfonyl-L- phenylalanyl)-L-lysine
The title compound was prepared from Nα-isobutyl-Λfø-(4-methylbenzenesulfonyl)-L-Iysine hydrochloride (120 mg, 0.34 mmol, example 1, step E) as described in general procedure Be using Nα-benzenesulfonyl-L-phenylalanine (300 mg, 1.0 mmol) prepared in step A of this example. The final product was purified by preparative HPLC to yield 107 mg (56%) of the desired material.
'H ΝMR (CDC13): δ 0.74 (d, J= 6.3, 3H), 0.80 (d, J= 6.3, 3H), 1.10-1.21 (m, 2H), 1.26-1.33 (m, 2H), 1 J0-1 J4 (m, 1H), 1.89-1.93 (m, 2H), 2.39 (s, 3H), 2.79-2.89 (m, 2H), 3.85 (t, J= 5.9, 1H), 4.29 (t, J= 6.9, 1H), 6.90 (d, J= 6.2, 2H), 7.08-7.29 (m, 6H), 7.35 (t, = 6.2, 2H), 7.44 (d, J= 8.1, 2H), 7.73 (d, J= 8.1, 2H). LC-MS: 642.2 (M - H)\ 99% pure. Example 9. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesuIfonyI)-Ne-[N'α-(4- chlorobenzenesulfonyl)-L-phenylalanyl]-L-lysine
Step A. Preparation of Nα-(4-chlorobenzenesulfonyl)-L-phenylalanine
L-phenylalanine was reacted with 4-chlorobenzenesulfonyl chloride under the conditions used in general procedure A giving the title compound which was recrystallised neat (30%).
LC-MS: 338 (M - H)\ 98% pure.
Step B. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-[N'α-(4- chlorobenzenesulfonyl)-L-phenylalanyl]-L-lysine
The title compound was prepared from Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine (100 mg, 0.29 mmol, example 1, step E) as described in general procedure Be using Na-(4- chlorobenzenesulfonyl)-L-phenylalanine (89 mg, 0.3 mmol) prepared in step A of this example. The final product was purified by preparative HPLC to yield 56 mg (33%) of the desired material.
'H ΝMR (DMSO-d6): δ 0.74 (d, J= 6.3, 3H), 0.80 (d, J= 6.3, 3H), 1.00-1.11 (m, 4H), 1.23-1.25 (m, IH), 1.70-1.74 (m, IH), 1.89-1.93 (m, IH), 2.32 (s, 3H), 2.59-2.67 (m, 2H), 2.84 and 2.95 (ABX, J= 14.1, 6.8, 2H), 3.85 (t, J= 5.9, IH), 4.11 (t, J= 6.9, IH), 7.02-7.21 (m, 7H), 7.24 (d, J= 8.0, IH), 7.54 (d, J= 8.1, 2H), 7.73 (d, J= 8.1, 2H), 8.07 (d, J= 6.4, IH). LC-MS: 677.2 (M - H)-, 99% pure.
Example 10. Preparation of Nα-isobutyl-Nα-(4-methyIbenzenesuIfonyl)-Ne-[N,α-(4- nitrobenzenesulfonyl)-L-phenylalanyl]-L-lysine
Step A. Preparation of Nα-(4-nitrobenzenesulfonyl)-L-phenylalanine L-phenylalanine was reacted with 4-nitrobenzenesulfonyl chloride under the conditions used in general procedure A giving the title compound which was recrystallised in ether (37%).
LC-MS: 349 (M - H)\ 98% pure.
Step B. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-[N'α-(4- nitrobenzenesulfonyl)-L-phenylalanyl]-L-lysine
The title compound was prepared fromNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine (100 mg, 0.29 mmol, example 1, step E) as described in general procedure Be using Nα-(4- nitrobenzenesulfonyl)-L-phenylalanine (125 mg, 0.5 mmol) prepared in step A of this example. The final product was purified by preparative HPLC to yield 91 mg (52%) of the desired material.
LC-MS: 687.2 (M - H)\ 98% pure.
Example 11. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-[N'α-(4- methylbenzenesulfonyl)-L-tyrosyl]-L-lysine
Step A. Preparation of Nα-(4-methylbenzenesulfonyl)-L-tyrosine
L-tyrosine was reacted with 4-methylbenzenesulfonyl chloride under the conditions used in general procedure A giving the title compound which was recrystallised neat (15%).
LC-MS: 334 (M - H)", 95% pure.
Step B. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-[N'α-(4- methylbenzenesulfonyl)-L-tyrosyl]-L-lysine
The title compound was prepared from Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine (100 mg, 0.29 mmol, example 1 , step E) as described in general procedure Be using Nα-(4- methylbenzenesulfonyl)-L-tyrosine (160 mg, 0.5 mmol) prepared in step A of this example. The final product was purified by preparative HPLC to yield 78 mg (46%) of the desired material.
LC-MS: 672.2 (M - H)\ 99% pure.
Example 12. Preparation of Nα-isobutyl-N -(4-methylbenzenesulfonyl)-Ne-[N,α-(4- a m in o b enzen es ulf o ny l)-L-try p toph a ny I] -L-lysin e
The title compound was prepared from Nα-isobutyl-Nα-(4-methylberιzenesulfonyl)-L-lysine hydrochloride (200 mg, 0.59 mmol, example 1 , step E) as described in general procedure Be using Nα-(4-nitrobenzenesulfonyl)-L-tryptophan (300 mg, 0.75 mmoϊ) prepared in step A of example 5. The intermediate, Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-[N' -(4- nitrobenzenesulfonyl)-L-tryptophanyl]-L-]ysine, was reduced following the conditions of general procedure E. The final product was purified by preparative HPLC to yield 266 mg (58%) of the • desired material.
Η ΝMR (DMSO-d6): δ 0.74 (d,J= 6.3, 3H), 0.80 (d,J= 6.3, 3H), 1.00-1.11 (m,4H), 1.32-1.35 (m, IH), 1.70-1.74 ( , IH), 1.83-1.88 (m, IH), 2.34 (s, 3H), 2.59-2.67 ( , 2H), 2.84-2.96 (m, 2H)r 3.86 (t, J= 6.5, IH), 4.13 (t, J= 7.2, IH), 6.85 (t, - 4.0, IH), 7.00 (t, J= 4.0, 2H), 7.10 (d, J= 7.1, 2H), 7.28 (d, J= 4.0, IH), 7.33 ( , 3H), 7.43 (d, J= 7.1, 2H), 7.60 (d, J= 7.0, 2H), 7.71 (t. J= 4.1 , IH), 7.80 (d, = 8.0, IH). LC-MS: 725.2 (M - H)\ 98% pure.
Example 13. Preparation of Λ'α-isobutyI-Nα-(4-methylbenzenesulfonyl)-Ne-[N'α-(4- nitrobenzenesuIfonyI)-L-aIanyl]-L-Iysine
Step A. Preparation of Nα-(4-nitrobenzenesulfonyl)-L-alanine
L-alanine was reacted with 4-nitrobenzenesulfonyI chloride under the conditions used in general procedure A giving the title compound which was recrystallised neat (9%). This compound was used without further purification in the next step. Step B. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-[N' -(4- nitrobenzenesulfonyl)-L-alanyl]-L-lysine
The title compound was prepared from Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine hydrochloride (100 mg, 0.29 mmol, example 1, step E) as described in general procedure Be using Nα-(4-nitrobenzenesulfonyl)-L-alanine (140 mg, 0.5 mmol) prepared in step A of this example. The final product was purified by preparative HPLC to yield 10 mg (6.5%) of the desired material.
LC-MS: 611.2 (M - H)\ 99% pure.
Example 14. Preparation of Nα-isobutyI-Nα-(4-methyIbenzenesulfonyl)-Ne-[N,α- (benzenesulfonyl)-L-non'alyl]-L-lysine
Step A. Preparation of Nα-benzenesulfonyl-L-norvaline
L-norvaline was reacted with benzenesulfonyl chloride under the conditions used in general procedure A giving the title compound which was recrystallised neat (33%). This compound was used without further purification in the next step.
Step B . Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-[N 'α-(benzenesulfonyl)- L-norvalyl]-L-lysine
The title compound was prepared from Nα-isobutyl-Ntx-(4-methylbenzenesulfonyl)-L-ly sine (100 mg, 0.29 mmol, example 1, step E) as described in general procedure Be using Nα- benzenesulfonyl-L-norvalinc (120 mg, 0.5 mmol) prepared in step A of this example. The final product was purified by preparative HPLC to yield 15 mg (10%) of the desired material.
LC-M S: 594.3 (M - H)\ 99% pure. Example 15. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-[N'α- (benzenesulfonyl)-L-norleucyl]-L-lysine
Step A. Preparation of N -benzenesulfonyl-L-norleucine
L-norleucine was reacted with benzenesulfonyl chloride under the conditions used in general procedure A giving the title compound which was recrystallised neat (25%). This compound was used without further purification in the next step.
Step B. Preparation ofNα-isobutyl-Λ7α-(4-methylbenzenesulfonyl)-Ne-[N' -(benzenesulfonyl)- L-norleucyl]-L-lysine
The title compound was prepared from Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine (100 mg. 0.29 mmol, example 1, step E) as described in general procedure Be using Nα- benzenesulfonyl-L-norleucine (125 mg, 0.5 mmol) prepared in step A of this example. The final product was purified by preparative HPLC to yield 13 mg (8.5%) of the desired material.
LC-MS: 608.3 (M - H)", 99% pure.
Example 16. Preparation of Nα-isoburj'l-Nα-(4-methyIbenzenesuIfonyI)-Ne-[N,α-(4- nitrobenzenesulfonyl)-L-leucyl]-L-lysine
Step A. Preparation of Nα-(4-nitrobenzenesulfonyl)-L-leucine
L-leucine was reacted with 4-nitrobenzenesulfonyl chloride under the conditions used in general procedure A giving the title compound which was recrystallised neat (66%). This compound was used without further purification in the next step.
Step B. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-[N'α-(4- nitrobenzenesulfonyl)-L-leucyl]-L-lysine The title compound was prepared fromNα-isobutyl-No!-(4-methylbenzenesulfonyl)-L-lysine (100 mg, 0.29 mmol, example 1, step E) as described in general procedure Be using Nα-(4- nitrobenzenesulfonyl)-L-leucine (150 mg, 0.5 mmol) prepared in step A of this example. The final product was purified by preparative HPLC to yield 28 mg (17%) of the desired material.
LC-MS: 653.3 (M - H)\ 99% pure.
Example 17. Preparation of Nα-isobutyl-Nα- .-rnethylbenzenesuIfonyrj-Ne-pN'ct- enzenesulfonyl^-trø/t.s^hydroxy-L-prolylJ-L-lysine
Step A. Preparation of Nα-benzenesulfonyl-4-trΛ«i,-hydroxy-L-proline
4-tz- π^-hydroxy-L-proline was reacted with benzenesulfonyl chloride under the conditions used in general procedure A giving the title compound which was recrystallised neat (21%). This compound was used without further purification in the next step.
Step B. Preparation ofΛ^ -isobutyl-Nα-(4-methyIbenzenesulfonyl)-Ne-[N'α-(benzenesulfonyl)- 4-tr-7/M-hydroxy-L-ρrolyl]-L-lysine
The title compound was prepared fromNα-isobutyl-7 α-(4-methylbenzenesulfonyl)-L-]ysine (100 mg, 0.29 mmol, example 1, step E) as described in general procedure Be using Nα- benzenesulfonyl-4-trøn.s'-hydroxy-L-proline (130 mg, 0.5 mmol) prepared in step A of this example. The final product was purified by preparative HPLC to yield 9 mg (6%) of the desired material.
LC-MS: 608.3 (M - H)\ 99% pure.
Example 18. Preparation of Nα-isobutyI-Nα-(4-methyIbenzenesuIfonyI)-Ne-[N'α-(4- fluorobenzenesulfonyl)-L-phenylalanyl]-L-lysine
Step A. Preparation of Nα-(4-fluorobenzenesulfonyl)-L-phenylalanine L-phenylalanine was reacted with 4-fluorobenzenesulfonyl chloride under the conditions used in general procedure A giving the title compound which was recrystallised in ether (40%).
LC-MS: 322 (M - H)\ 99% pure.
Step B. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-[N' -(4- fluorobenzenesulfonyl)-L-phenylalanyl]-L-lysine
The title compound was prepared fromNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine (100 mg, 0.29 mmol, example 1, step E) as described in general procedure Be using N -(4- fluorobenzenesulfonyl)-L-phenylalanine (160 mg, 0.5 mmol) prepared in step A of this example. The final product was purified by preparative HPLC to yield 87 mg (52%) of the desired material.
!H ΝMR (DMSO-d6): δ 0.74 (d,/= 6.3, 3H), 0.80 (d, = 6.3, 3H), 1.00-1.11 (m,4H), 1.23-1.25 (m, IH), 1.70-1.74 (m, IH), 1.89-1.93 (m, IH), 2.32 (s, 3H), 2.65 (m, 2H), 2.85 and 2.95 (ABX, J= 16.1 7.1, 2H), 3.88 (t, J= 6.0, IH), 4.11 (t, .7= 6.9, IH), 7.02-7.21 (m, 7H), 7.24 (d, J= 8.0, IH), 7.54 (d, J= 8.1, 2H), 7.73 (d, J= 8.1, 2H), 8.15 (d, J= 6.6, IH). LC-MS: 660.2 (M - H)\
99% pure.
Example 19. Preparation of Λrα-isobutyl-No:-(4-methyIbenzenesulfonyl)-Ne-[N,α-(2- naphthylsulfonyl)-L-phenylalanyl]-L-lysine
Step A. Preparation of Nα-(2-naphthylsulfonyl)-L-phenylalanine
L-phenylalanine was reacted with 2-naphthylsulfonyl chloride under the conditions used in genera] procedure A giving the title compound which was recrystallised in ether (51 %).
LC-MS: 352 (M - H)\ 95% pure.
Step B. Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-[N'α-(2-naphthylsulfony3)- L-phenylalanyl]-L-lysine The title compound was prepared from Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-ly sine (100 mg, 0.29 mmol, example 1, step E) as described in general procedure Be using Nα-(2- naphfhylsulfonyl)-L-phenylalanine (175 mg, 0.5 mmol) prepared in step A of this example. The final product was purified by preparative HPLC to yield 55 mg (32%) of the desired material.
LC-MS: 692.3 (M - H)\ 99% pure.
Example 20. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesuIfonyI)-Ne-[N'α-(4- bromobenzenesulfonyl)-L-phenylalanyl]-L-lysine
Step A. Preparation of Nα-(4-bromobenzenesulfonyl)-L-phenylalanine
L-phenylalanine was reacted with 4-bromobenzenesulfonyl chloride under the conditions used in general procedure A giving the title compound which was recrystallised in ether (18%).
LC-MS : 383 (M - H)\ 99% pure.
Step B. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-[N'α-(4- bromobenzenesulfonyl)-L-phenylalanyl]-L-lysine
The title compound was prepared from Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-ly sine (100 mg, 0.29 mmol, example 1, step E) as described in general procedure Be using Nα-(4- bromobenzenesulfonyl)-L-phenylalanine (190 mg, 0.5 mmol) prepared in step A of this example. The final product was purified by preparative HPLC to yield 56 mg (30%) of the desired material.
,H ΝMR (DMSO-d6): δ 0.74 (d, = 6.3, 3H), 0.80 (d,J= 6.3, 3H), 1.00-1.11 (m, 4H), 1.23-1.25 (m, IH), 1.70-1.74 (m, IH), 1.89-1.93 (m, IH), 2.32 (s, 3H), 2.60-2.68 (m, 2H), 2.86 and 2.97(ABX, J= 14.1, 7.1, 2H), 3.96 (t, J= 6.1, 1H), 4.11 (t,.7= 6.9, IH), 6.99-7.22 (m, 7H), 7.24 (d, J= 8.0, IH), 7.54 (d, J= 8.1, 2H), 7.73 (d, J= 8.1, 2H), 8.11 (d, J= 6.5, IH). LC-MS: 721.2 (M - H)-, 99% pure. Example 21. Preparation of Nα-isobutyI-Nα-(4-methyIbenzenesuIfonyI)-Ne-[NV(4- methylbenzenesulfonyl)-glycyl]-L~lysine
Step A. Preparation of Nα-(4-methylbenzenesulfonyl)-glycine
Glycine was reacted with 4-methylbenzenesulfonyl chloride under the conditions used in general procedure A giving the title compound which was recrystallised neat (60%).
LC-MS: 228 (M - H)\ 99% pure.
Step B. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-(9- fluorenylmethoxycarbonyl)-L-lysine
The title compound was prepared from Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine by following the indications of general example Be using commercially available N-(9- fluorenylmethoxycarbonyloxy) succinimide instead of the usual combination of reactants i.e. N- protected amino acid, N-hydroxysuccinimide and DCC.
Η ΝMR (CDC13): 0.79 (d, J= 7.1, 3H), 0.81 (d, J= 7.1 , 3H), 1.12-1.25 (m, 2H), 1.30-1.40 (m, 2H), 1.42-1.50 (m, 2H), 1.78-1.90 (m, 2H), 2.36 (s, 3H), 2.85 (m, 2H), 2.88 and 3.04 (ABX, J = 14.3, 7.3, 2H), 4.16-4.21 (m, 2H), 4.28 (d, J= 7.0, 2H), 7.30-7.42 (m, 6H), 7.60 (m, 4H), 7.88 (d, J = 7.5, 2H), 12.69 (br s. lH).
Step C. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-[N'α-(4- methylbenzenesulfonyl)-glycyl]-L-lysine
The title compound was prepared from solid phase bound Nα-isobutyl-Nα-(4- methylbenzenesulfonyl)-Ne-(9-fluorenylmethoxycarbonyl)-L-lysine (step B) as described in general procedure Bb using Nα-(4-methylbenzenesulfonyl)-glycine (110 mg, 0.5 mmol) prepared in step A of this example. The final product was purified by preparative HPLC to yield 30 mg (42%) of the desired material.
Η NMR CCDCla): δ 0.73 (d, J= 6.9, 6H), 1.23-1.25 (m, 2H), 1.45-1.52 (m, 3H), 1.89-1.99 (m, 2H), 2.32 (s, 6H), 2.94-3.03 (m, 2H)5 3.16 (m, 2H), 3.55 (t, J= 5.9, IH), 4.27 (t, J=. 7.2, IH), 7.26 (d, J= 8.1, 4H), 7.73 (d, J= 8.1, 4H). LC-MS: 566.5 (M - H)\ 85% pure.
Example 22. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-(N'α- benzenesulfonyl-L-leucyl]-L-lysine
Step A. Preparation of Nα-benzenesulfonyl-L-leucine
L-leucine was reacted with benzenesulfonyl chloride under the conditions used in general procedure A giving the title compound which was recrystallised neat (31 %). This compound was used without further purification in the next step.
Step B. Preparation of Na-isobutyl-Na-(4-methylbenzenesulfonyl)-Ne-(Ar'a-benzenesulfonyl-L- leucyl]-L-lysine
The title compound was prepared from solid phase bound 7 α-isobutyl-Nα-(4- methylbenzenesulfonyl)-Ne-(9-fluorenylmethoxycarbonyl)-L-lysine as described in general procedure Bb using Nα-benzenesulfonyl-L-leucine (130 mg, 0.5 mmol) prepared in step A of this example. The final product was purified by preparative HPLC to yield 30 mg (39%) of the desired material.
Η ΝMR (CDC13): δ 0.72 (d, = 6.0, 3H), 0.75 (d, J= 6.0, 3H), 0.78-0.81 (m, 6H), 1.20-1.22 (m, 2H)S 1.32-1.34 (m, 2H), 1.52-1.55 (m, IH), 1.78-2.04 (m, 3H), 2.32 (s, 3H), 2.81-3.01 (m, 4H), 3.56 (t, J= 5.2, IH), 4.25 (t, J= 6.0, IH), 7.21-7.29 (m, 2H), 7.42-7.45 (m, 2H), 7.52 (t, .7= 6.1, IH), 7.71 (d, J= 7.8, 2H), 7.81 (d, J= 7.8, 2H). LC-MS: 608.2 (M - H)\ 90% pure. Example 23. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-[N,α-(4- trifluoromethylbenzenesulfonyl)-L-phenylalanyl]-L-lysine
Step A. Preparation of Nα-(4-trifluoromethylbenzenesulfonyl)-L-phenylalanine
L-phenylalanine was reacted with 4-trifluoromethylbenzenesulfonyl chloride under the conditions used in general procedure A giving the title compound which was recrystallised neat (18%>).
LC-MS: 369 (M - H)\ 98% pure.
Step B. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-[N'α-(4- trifluoromethylbenzenesulfonyl)-L-phenylalanyl]-L-lysine
The title compound was prepared from solid phase bound Nα-isobutyl-Nα-(4- methylbenzenesulfonyl)-Ne-(9-fluorenyImethoxycarbonyl)-L-lysine as described in general procedure Bb using Nα-(4-trifluoromethylbenzenesulfonyl)-L-phenylalanine (180 mg, 0.5 mmol) prepared in step A of this example. The final product was purified by preparative HPLC to yield 50 mg (56%) of the desired material.
LC-MS: 710.2 (M - H)", 80% pure.
Example 24. Preparation of Nα-isoburyl-Nα-(4-rnethylbenzenesulfonyl)-Ne-[N'α-(2- thiophenesulfonyl)-L-phenylalanyl]-L-lysine
Step A. Preparation of Nα-(2-thiophenesulfonyl)-L-phenylalanine
L-phenylalanine was reacted with 2-thiophenesulfonyl chloride under the conditions used in genera] procedure A giving the title compound which was recrystallised from ether (93%).
LC-MS: 310 (M - H)\ 99% pure. Step B. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-[N'α-(2- thiophenesulfonyl)-L-phenylalanyl]-L-lysine
The title compound was prepared from Nα-isobuty'l-Nα-(4-methylbenzenesulfonyl)-L-lysine hydrochloride (250 mg, 0.61 mmol, example 1, step E) as described in general procedure Be using Nα-(2-thiophenesulfonyl)-L-phenylalarιine (155 mg, 0.5 mmol) which was prepared in step A of this example. The crude material was purified by HPLC to give 272 mg (66%) of pure adduct.
Η ΝMR (DMSO-d6): δ 0.77 (d,J= 6.3, 3H), 0.8 (d,J= 6.3, 3H), 1.00-1.11 (m, 4H), 1.32-1.35 (m, IH), 1J0-1J4 (m, IH), 1.83-1.88 (m, IH), 2.34 (s, 3H), 2.59-2.67 (m, 2H), 2.92 and 2.96 (ABX, J= 16.5, 7.1, 2H), 3.90 (t, J= 6.5, IH), 4.11 (t, J= 7.2, IH), 7.00 (t, J= 4.0, IH), 7.10- 7.21 (m, 5H), 7.30 (d, J= 4.0, IH), 7.34 (d, J= 8.1, 2H), 7.63 (d, .7= 8.1, 2H), 7.77 (d, J= 4.0, IH), 7.81 (t, J = 5.3, IH), 8.22 (d, J= 8.1, IH). LC-MS: 648.5 (M - H)\ 99% pure.
Example 25. Preparation of Nα-isobutyl-Λrα-(4-methylbenzenesulfonyl)-Ne-(N'α- benzenesuIfonyI)-L-asparagyl]-L-lysine
Step A. Preparation of Nα-benzenesulfonyl-L-asparagine
L-asparagine was reacted with benzenesulfonyl chloride under the conditions used in general procedure A giving the title compound which was recrystallised neat (29%). This compound was used without further purification in the next step.
Step B. Preparation ofN-t-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-(N'α-benzenesulfonyl)-L- asparagyl]-L-lysine
The title compound was prepared from solid phase bound Nα-isobutyl-Nα-(4- methylbenzenesulfonyl)-Ne-(9-fluorenylmethoxycarbonyl)-L-lysine as described in general procedure Bb using Nα-benzenesulfonyl-L-asparagine (350 mg, 1.0 mmol) prepared in step A of this example. The final product was purified by preparative HPLC to yield 70 mg (91%) of the desired material.
ΗNMR (CDC13): δ 0.78 (d, J= 6.3, 3H), 0.81 (d, J= 6.3, 3H), 0.94-1.13 (m, 4H), 1.32-1.35 (m, IH), 1.66-1.69 (m, IH), 1.83-1.88 (m, IH), 2.10-2.29 (m, 2H), 2.35 (s, 3H), 2.38 (s, 3H), 2.82 and 2.99 (ABX, J= 12.6, 8.1, 2H)5 4.00 (t,
Figure imgf000068_0001
7.9, 2H), 7.32 (d, J= 7.9, 2H), 7.55-7.64 (m, 3H), 7.77 (d, J= 7.8, 2H). LC-MS: 609.1 (M - H)", 99% pure.
Example 26. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-[N'α-(4- methylbenzenesulfonyl)-L-4-nitrophenylalanyl]-L-lysine
Step A. Preparation of Nα-(4-methylbenzenesulfonyl)-L-4-nitrophenyla]anine
L-4-nitrophenyl alanine was reacted with 4-methylbenzenesulfonyl chloride under the conditions used in general procedure A giving the title compound which was recrystallised neat (14%).
LC-MS: 364 (M - H)", 98% pure.
Step B. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-[N'α-(4- methylbenzenesuIfonyI)-L-4-nitrophenylaIanyl]-L-Iysine
The title compound was prepared from solid phase bound Nα-isobutyI-Nα-(4- methylbenzenesulfonyl)-Ne-(9-fluorenylmethoxycarbonyl)-L-lysine as described in general procedure Bb using Nα-(4-methylbenzenesulfonyl)-L-4-nitrophenylalanine (180 mg, 0.5 mmol) prepared in step A of this example. The final product was purified by preparative HPLC to yield 25 mg (28%) of the desired material.
LC-MS: 701.1 (M - H)", 95% pure. Example 27. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-[N'α-(4- methylbenzenesulfonyl)-L-phenylglycyl]-L-lysine
Step A. Preparation of Nα-(4-methylbenzenesulfonyl)-L-phenylglycine
L-phenylglycine was reacted with 4-methylbenzenesulfonyl chloride under the conditions used in general procedure A giving the title compound which was recrystallised neat (21%).
LC-MS: 288 (M - H)", 99% pure.
Step B. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-[N'α-(4- methylbenzenesulfonyl)-L-phenylglycyl]-L-lysine
The title compound was prepared from solid phase bound Nα-isobutyl-Nα-(4- methylbenzenesulfonyl)-Ne-(9-fluorenylmethoxycarbonyl)-L-lysine as described in general procedure Bb using Nα-(4-methylbenzenesulfonyl)-L-phenylglycine (150 mg, 0.5 mmol) prepared in step A of this example. The final product was purified by preparative HPLC to yield 55 mg (68%) of the desired material.
LC-MS: 642.1 (M - H)', 95% pure.
Example 28. Preparation of Nα-isobutyl-Nα-(4-nitrobenzenesuIfonyI)-Ne-[N'α-(4- acetamidobenzenesulfonyl)-L-phenylalanyl]-L-lysine
Step A. Preparation of Nα-isobutyl-Nα-(4-nitrobenzenesulfonyl)-Ne-(9- fluorenylmethoxycarbonyl)-L-lysine
Λfø-isobutyl-Nα-(4-nitrobeιιzenesulfonyl)-L-]ysine hydrochloride (1 mmol, example 5, step C) was partially dissolved in K2CO3 (1 M)/THF/CH3CΝ (4 mL/4 mL/4 mL). To this suspension was added N-(9-fluorenylmethoxycarbonyloxy) succinimide (371 mg, 1.10 mmol). The reaction turned slowly to colorless and was left stirring for 1 h. HCl (IM) was added until acidic pH and the reaction mixture was extracted twice with EtOAc. The combined organic layers were washed with brine, dried over MgSO4 and concentrated. The residue was purified by flash chromatography eluting with a mixture of hexane/EtOAc containing 0.4% AcOH to yield 88% of the title compound which was used without further purification in the next step.
Step B. Preparation of Nα-isobutyl-Nα-(4-nitrobenzenesulfonyl)-Ne-[N'α-(4- acetamidobenzenesulfonyl)-L-phenylalanyl]-L-lysine
The title compound was prepared from solid phase bound Nα-isobutyl-Nα-(4- nitrobenzenesulfonyl)-Ne-(9-fluorenylmethoxycarbonyl)-L-lysine (step A) as described in general procedure Bb using Nα-(4-acetamidobenzenesulfonyl)-L-phenylalanine (400 mg, 1.2 mmol) which was prepared in step A of example 3. The final product was purified by preparative HPLC to yield 55 mg (60%) of the desired material.
LC-MS: 730.1 (M - H)", 95% pure.
Example 29. Preparation of Λrα-isobutyl-Nα-(4-nitrobenzenesulfonyl)-Ne-[N'α-(2- thiophenesulfonyι)-L-phenylalanyl]-L-lysine
The title compound was prepared from solid phase bound Nα-isobutyl-Nα-(4- nitrobenzenesulfonyl)-Ne-(9-fluorenylmethoxycarbonyl)-L-lysine as described in general procedure Bb using Nα-(2-thiophenesulfonyl)-L-phenylalanine (300 mg, 1.2 mmol) which was prepared in step A of example 24. The final product was purified by preparative HPLC to yield 46 mg (54%o) of the desired material.
LC-MS: 679.0 (M - H)\ 95% pure.
Example 30. Preparation of Nα-isobu 'I-Nα-(4-nιethylbenzenesulfonyl)-Ne-(N'α-acetyl-L- phenylalanyl)-L-lysine
Step A. Preparation of Nα-acetyl-L-phenylalanine L-phenylalanine was reacted with acetyl chloride under the conditions used in general procedure A giving the title compound which was recrystallised from ether (97%>). This compound is also commercially available.
LC-MS : 206 (M - H)\ 99% pure.
Step B. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-(N'α-acetyl-L- phenylalanyl)-L-lysine
The title compound was prepared from Nα-isobμtyd-Nα-(4-methylbenzenesulfonyl)-L-lysine (150 mg, 0.42 mmol, example 1, step E) as described in general procedure Be using Nα-acetyl-L- phe ylalanine (207 mg, 1.0 mmol) which was prepared in step A of this example. The final product was purified by preparative HPLC'to yield 121 mg (59%) of the desired material.
ΗΝMR (CDC13): δ 0.83 (d, 7= 6.9, 6H), 1.08-1.1 l (m, 2H), 1.23-1.25 (m, 2H), 1.45-1.52 (m, IH), 1.89-1.99 (m and s (1.90), 5H), 2.32 (s, 3H), 2.94-3.09 (m, 6H), 4.23 (t, IH J= 5.9), 4.61 (m, IH), 7.09-7.26 (m, 7H), 7.73 (d, J= 8.1, 2H). LC-MS: 544.2 (M - H)\ 99% pure.
Example 31. Preparation of Nα-isobutyl-Λrα-(4-methylbenzenesuIfonyl)-Ne-(Λr'α- benzyloxycarbonyI-L-phenylaIanyI)-L-lysine
The title compound was prepared from solid phase bound Nα-isobutyl-Nα-(4- methylbenzenesulfonyl)-Ne-(9-fluorenylmethoxycarbonyl)-L-lysine as described in general procedure Bb using commercially available Nα-benzyloxy carbonyl -L-phenylalanine (300 mg, 1.2 mmol). The final product was purified by preparative HPLC to yield 33 mg (37%) of the desired material.
LC-MS: 636.2 (M - H)" s 99% pure. Example 32. Preparation of Nα-isobutyI-Nα-(4-methylbenzenesulfonyl)-Ne-[N'α-(4- methylbenzenesulfonyl)-L-seryl)-L-lysine
Step A. Preparation of Nα-(4-methylbenzenesulfonyI)-L-serine
L-serine was reacted with 4-methylbenzenesulfonyl chloride under the conditions used in general procedure A giving the title compound which was recrystallised neat (44%). This compound was used without further purification in the next step.
Step B. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-[N'α-(4- methyIbenzenesulfonyl)-L-seryl]-L-lysine
The title compound was prepared from solid phase bound Nα-isobutyl-Nα-(4- methy]benzenesulfonyl)-Ne-(9-fluorenylmethoxycarbonyl)-L-lysine as described in general procedure Bb using Nα-(4-mclhyIbcnzcncsulfonyl)-L-scrine (150 mg, 1.2 mmol) prepared in step A of this example. The final product was purified by preparative HPLC to yield 35 mg (46%) of the desired material.
Η ΝMR CCDCla): δ 0.78-0.82 (m, 6H), 1.1.8-1.22 (m, 2H), 1.38-1.41 (m, 2H), 1.50-1.52 (m, IH), 1.79-1.96 (m, 2H), 2.32 (s, 6H), 2.85-2.97 (m, 2H), 3.06-3.19 (m, 2H), 3.82 (br s, IH), 4.23 (t; J= 6.9, IH), 7.25 (d, J= 8.0, 4H), 7.70 (d, J= 8.0, 4H). LC-MS: 596.1 (M - H)\ 95% pure.
Example 33. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-[N,α-(4- methylbenzenesulfonyl)-L-cyc]ohexy]alanyl]-L-lysine
Step A. Preparation of Nα-(4-methylbenzenesulfonyl)-L-cyclohexylalanine
L-cyclohexyl alanine was reacted with 4-methylbenzenesulfonyl chloride under the conditions used in general procedure A giving the title compound which was recrystallised neat (14%>).
LC-MS: 324 (M - H)', 95% pure. Step B. Preparation of Nα-isobutyl-Nα-(4-methylbcnzenesulfonyl)-Ne-[N'α-(4- methylbenzenesulfonyl)-L-cyclohexylalanyl]-L-lysine
The title compound was prepared from solid phase bound Nα-isobutyl-Nα-(4- methylbenzenesulfonyl)-Ne-(9-fluorenylmethoxycarbonyl)-L-lysine as described in general procedure Bb using Nα-(4-methyIbenzenesulfonyl)-L-cyclohexylalanine (350 mg, 1.2 mmOl) prepared in step A of this example. The final product was purified by preparative HPLC to yield 19 mg (22%o) of the desired material.
]H ΝMR (CDC13): δ 0.73-1.73 (m, 24H), 1.89-1.99 (m, 2H), 2.39 (s, 6H), 2.94-3.19 (m, 4H), 3.65 (t, J= 5.9, IH), 4.33 (t, J= 7.2, IH), 7.26 (d, = 8.1, 4H), 7.73 (d, = 8.1, 4H). LC-MS: 662.2 (M - H)\ 95% pure.
Example 34. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Λre-[Λr,α-(4- methylbenzenesulfonyl)-L-glutaminyl]-L-lysine
Step A. Preparation of Nα-(4-methylbenzenesulfonyl)-L-glutamine
L-glutamine was reacted with 4-methylbenzenesulfonyl chloride under the conditions used in general procedure A giving the title compound which was recrystallised neat (11%). This compound was used without further purification in the next step.
Step B. Preparation of • Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-[N'α-(4- methylbenzenesulfonyl)-L-glutaminyl]-L-lysine
The title compound was prepared from solid phase bound Nα-isobutyl-Nα-(4- methylbenzenesulfonyl)-Ne-(9-fluorenylmethoxycarbonyl)-L-lysine as described in general procedure Bb using Nα-(4-methylbenzenesulfonyl)-L-glutamine (360 mg, 1.2 mmol) prepared in step A of this example. The final product was purified, by preparative HPLC to yield 18 mg (21%) of the desired material. LC-MS: 637.2 (M - H)\ 80% pure.
Example 35. Preparation of Nα-isobutyI-Nα-(4-methyIbenzenesulfonyl)-Ne-[N'α-(4- methyIbenzenesuIfonyl)-L-2-thiophenyIaIanyI]-L-lysine
Step A. Preparation of Nα-(4-methylbenzenesulfonyl)-L-2-thiophenylalanine
L-2-thiophenylalanine was reacted with 4-methylbenzenesulfonyl chloride under the conditions used in general procedure A giving the title compound which was recrystallised neat (33 %). This compound was used without further purification in the next step.
Step B. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-[N'α-(4- methylbenzenesulfonyl)-L-2-thiophenylalanyl]-L-lysine
The title compound was prepared from solid phase bound Nα-isobutyl-Nα-(4- methylbenzenesulfonyl)-Ne-(9-fluorenylmethoxycarbonyl)-L-lysine as described in general procedure Bb using Nα-(4-methylbe:nzenesulfonyl)-L-2-thiophenylalanine (390 mg, 1.2 mmol) prepared in step A of this example. The final product was purified by preparative HPLC to yield 15 mg (18%) of the desired material.
LC-MS: 662.1 (M - H)\ 85% pure.
Example 36. Preparation of Nα-isobutyl-Arα-(4-methyIbenzenesuIfonyI)-Ne-[N,α-(9- fluorenylmethoxycarbonyl)-L-seryl]-L-lysine
The title compound was prepared from solid phase bound Nα-isobutyl-Nα-(4- methylbenzenesulfonyl)-Ne-(9-fluorenylmethoxycarbonyl)-L-lysine as described in general procedure Bb using commercially available Nα-(9-fluorenylmethoxycarbonyl)-L-serine (390 mg, 1.2 mmol). The final product was purified by preparative HPLC to yield 15 mg (18%) of the desired material. LC-MS: 664.3 (M - H)\ 95% pure.
Example 37. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-[Λr'α-(9- fluorenylmethoxycarbonyl)-L-cyclohexylalanyl]-L-lysine
The title compound was prepared from solid phase bound Nα-isobutyl-Nα-(4- methylbenzenesulfonyl)-Ne-(9-fluorenylmethoxycarbonyl)-L-lysine as described in general procedure Bb using commercially available Nα-(9-fluorenylmethoxycarbonyl)rL- cyclohexylalanine (470 mg, 1.2 mmol). The final product was purified by preparative HPLC to yield 11 mg (12%) of the desired material.
LC-MS: 730.6 (M - H)\ 95% pure.
Example 38. Preparation of Nct-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-[N'α-(4- methylbenzenesulfonyl)-L-glutamyl]-L-lysine
Step A. Preparation of Nα-(4-methylbenzenesulfonyl)-L-glutamic acid
L-glutamic acid was reacted with 4-methylbenzenesulfonyϊ chloride under the conditions used in general procedure A giving the title derivative (20%) which was used without purification in the next step.
Step B. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-[N'α-(4- methylbenzenesulfonyl)-L-glutamyl]-L-lysine
The title compound was prepared from solid phase bound Nα-isobutyl-Nα-(4- methylbenzenesulfonyl)-Λ7e-(9-fluorenylmethoxycarbonyl)-L-lysine as described in general procedure Bb using Nα-(4-methylbenzenesulfonyl)-L-glutamic acid (360 mg, 1.2 mmol) prepared in step A of this example. The final product was purified by preparative HPLC to yield 20 mg (25%) of the desired material. LC-MS: 640.3 (M + H)+, 99% pure.
Example 39. Preparation of Nα-isobutyI-Nα-(4-methyIbenzenesuIfonyI)-Ne-[N,α-(4- methylbenzenesulfonyl)-L-lysyl]-L-lysine
Step A. Preparation of Nα-(4-methylbenzenesulfonyl)-Ne-tert-butoxycarbonyl-L-Iysine
Ne-terr-butoxycarbonyl-L-lysine was reacted with 4-methylbenzenesulfonyl chloride under the conditions used in general procedure A giving the title derivative which was used without purification in the next step.
Step B. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-[N'α-(4- methylbenzenesulfonyl)-L-lysyl]-L-lysine
The title compound was prepared from solid phase bound Nα-isobutyl-Nα-(4- methylbenzenesulfonyl)-Ne-(9-fluorenylmethoxycarbonyl)-L-lysine as described in general procedure Bb using Nα-(4-methylbenzenesulfonyl)-Ne-te7"/-butoxycarbonyl-L-lysine (360 mg, 1.2 mmol) prepared in step A of this example. The final product was purified by preparative HPLC to yield 10 mg (12%) of the desired material.
LC-MS: 637.2 (M - H)\ 99% pure.
Example 40. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Λre-[N'α-(4- methylbenzenesulfonyl)-O-benzyl-L-seryl]-L-lysine
The title compound was prepared from solid phase bound Nα-isobutyl-Nα-(4- methylbenzenesulfonyl)-Ne-(9-fluorenylmethoxycarbonyl)-L-lysine (200 mg, 0.1 mmol) in a similar fashion to general procedure Bb using commercially available Nα-(9- fluorenylmethoxycarbonyl)-O-benzyl-L-serine (100 mg, 0.24 mmol) with DCC (100 mg, 0.48 mmol) and HOBt (50 mg, 0.37 mmol) as the activating reagents. The intermediate adduct was again deprotected and coupled with 4-methylbenzenesulfonyl chloride in CH2C12 before being cleaved from the resin with TFA. The final product was purified by preparative HPLC to yield 30 mg (42%)) of the desired material.
1HNMR (DMSO-d6): δ 0.78 (d, J= 6.5, 3H), 0.82 (d, J= 6.6, 3H), 1.15-1.45 (m, 5H), 1.84 (m, IH), 1.95 (m, IH), 2.34 (s, 3H), 2.36 (s, 3H), 2.79 (m, 2H), 2.91 (m, 2H), 3.42 (t, J= 5.7, 2H), 3.92 (t, J= 6.1, IH), 4.04 (m, IH), 4.37 (d, J= 5.7, 2H), 7.21 (d, J= 7.3, 2H), 7.23-7.33 (m, 7H), 7.65 (d, J= 8.2, 2H), 7.71 (d, J= 7.7, 2H), 7.93 (t, J « 4.0, IH), 7.95 (br s, IH). LC-MS: 688 (M + H)", 99% pure.
Example 41. Preparation of Nα-isobutyI-Nα-(4-methylbenzenesulfonyl)-Ne-[N'α-(4- methylbenzenesulfonyl)~L-aspartyl]-L~lysine
Step A. Preparation of Nα-(4-methylbenzenesulfonyl)-L-aspartic acid
L-aspartic acid was reacted with 4-methylbenzenesulfonyl chloride under the conditions used in general procedure A giving the title derivative (40%) which was used without purification in the next step.
Step B. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-[N'α-(4- methylbenzenesulfonyl)-L-aspartyl]-L-lysme
The title compound was prepared from solid phase bound Nα-isobutyl-Nα-(4- methylbenzenesulfonyl)-Λ7e-(9-fluorenylmethoxycarbonyl)-L-lysine as described in general procedure Bb using Nα-(4-methylbenzenesulfonyl)-L-aspartic acid (340 mg, 1.2 mmol) prepared in step A of this example. The final product was purified by preparative HPLC to yield 20 mg (26%>) of the desired material.
LC-MS: 624.1 (M - H)\ 99% pure. Example 42. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-[N'α-(4- methyIbenzenesuIfonyI)-L-3-(2-thianaphthyI)-aIanyI]-L-Iysine
Step A. Preparation of 4-methylbenzenesulfonyl-L-3-(2-thianaphthyl)-alanine
L-3-(2-thianaphthyl)-alanine was reacted with 4-methylbenzenesulfonyl chloride under the conditions used in general procedure A giving the title derivative which was recrystallised neat (34%). This compound was used without further purification in the next step.
Step B. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-[N'α-(4- methylbenzenesulfonyl)-L-3-(2-thianaphthyl)-alanyl]-L-lysine
The title compound was prepared from solid phase bound Nα-isobutyl-Nα-(4- methylbenzenesulfonyl)-Ne-(9-fluorenylmethoxycarbonyl)-L-lysine as described in general procedure Bb using 4-methylbenzenesulfonyl-L-3-(2-thianaphthyl)-alanine (450 mg, 1.2 mmol) prepared in step A of this example. The final product was purified by preparative HPLC to ield 25 mg (28%) of the desired material.
JH ΝMR (CDC13): δ 0.85-0.89 (m, 6H), 1.08-1.15 (m, 2H), 1.32-1.36 (m, 2H), 1.50-1.53 (m, 2H), 1.85-1.89 (m, 2H), 2.24 (s, 3H), 2.96-3.12 (m, 4H), 3.84-3.86 (m, IH), 4.22 (t, J= 5.4, IH), 6.90 (d, J= 6.8, 2H), 7.06 (s, IH), 7.19-7.28 (m, 8H), 7.46 (d, J= 6.8, IH), 7.74 (d, J= 6.8, 2H). LC-MS: 714.2 (M + H)+, 99% pure.
Example 43. Preparation of Λrα-(4-aminobenzenesuIfonyl)-Nα-isobutyl-Ne-[N'α-(4- methylbenzenesulfonyl)-L-tryptophanyl]-L-lysine
The title compound was prepared from Nα-isobutyl-Nα-(4-nitrobenzenesulfonyl)-L-lysine hydrochloride (350 mg, 1.0 mmol, example 5, step C) as described in general procedure Be using Nα-(4-methylbenzenesulfonyl)-L-tryptophan (335 mg, 1.1 mmol) which was prepared in step A of example 2. The final product was purified by preparative HPLC to yield 650 mg (90%) of Nα- (4-nitrobenzenesulfonyl)-Nα-isobutyl-Ne-[Λ7'α-(4-methylbenzenesulfonyl)-L-tryptophanyl]-L- lysine. The latter derivative (300 mg) was hydrogenolysed following the indications of general procedure E. Purification by HPLC gave the desired material (235 mg, 75%).
]H NMR (DMSO-d6): δ 0.74 (d,J= 6.3, 3H), 0.80 (d, J= 6.3, 3H), 1.00-1.11 (m, 4H), 1.32-1.35 (m, IH), 1.70-1.74 (m, IH), 1.83-1.88 (m, IH), 2.34 (s, 3H), 2.59-2.67 (m, 2H), 2.84-2.96 (m, 2H), 3.80 (t, J= 6.5, IH), 4.11 (t, J= 7.2, IH), 6.85 (t, J= 5.2, IH), 7.00 (t, J= 4.0, 2H), 7.10 (d, J= 5.3, 2H), 7.28 (d, J= 4.0, IH), 7.33 (m, 3H), 7.43 (d, J= 6.9, 2H), 7.60 (d, J= 6.9, 2H), 7.71 (t, J= 6.9, IH), 7.80 (d, J= 8.0, IH). LC-MS: 696.8 (M - H)\ 99% pure.
Example 44. Preparation of Arα-(4-aminobenzenesulfonyl)-Nα-isobutyl-Ne-(N'α- benzenesulfonyl-L-phenylalanyl)-L-lysine
The title compound was prepared from Nα-isobutyl-Nα-(4-nitrobenzenesulfonyl)-L-lysine (385 mg, 1.0 mmol, example 5, step C) as described in general procedure Be using Nα- benzenesulfonyl-L-phenylalanine (335 mg, 1.1 mmol) which was prepared in step A of example 8. The final product was purified by preparative HPLC to yield 550 mg (85%) of Nα-(4- nitrobenzenesulfonyl)-Nα-isobutyl-Ne-(N'α-benzenesulfonyl)-L-phenylalanyl)-L-lysine. The latter derivative (200 mg) was hydrogenolysed following the indications of general procedure E. Purification by HPLC gave the desired material (129 mg, 65%).
Η ΝMR (CDC13): δ 0.75 (d, J= 7.1, 3H), 0.88 (d, J= 7.1, 3H), 1.0-1.1 (m, 2H), 1.16-1.23 (m, IH), 1.56-1.58 (m, IH), 1.72-1.73 (m, IH), 1.82-1.91 (m, IH), 2.79-2.96 (m, 2H), 3.86 7 (t, = 6.2, IH), 4.09 (t, .7= 6.1, IH), 5.59 (s, IH), 6.55 (d, J= 7.5, 2H), 7.05 (d, J= 7.1, 2H), 7.11-7.19 (m, 4H), 7.38 (d, J= 6.9, 2H), 7.42 (t, J= 7.1, 2H), 7.51 (d, J= 7.1, 2H), 8.01 (d, J= 7.1, IH). LC-MS: 643.2 (M - H)\ 99% pure.
Example 45. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-[N'α-(4- methylbenzeήesulfonyl)-L-tryptophanyl]-L-lysine2,3-dihydroxypropyl ester
A solution of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-[N'α-(4-methylbenzenesulfonyl)-L- tryptophanyl]-L-lysine (product of example 2, 70 mg, 0.1 mmol) in DMF (1 mL) was treated with glycerol (100 mg) and EDC (100 mg, 0.5 mmol) and stirred overnight. The solution is then poured in 5% citric acid and extracted with EtOAc (5 mL). The solvent was evaporated and the residue was purified by preparative HPLC to yield 30 mg (40%>) of the desired ester.
LC-MS: 769.3 (M - H)\ 99% pure.
Example 46. Preparation of Nα-(4-aminobenzenesulfonyl)-Nα-isobutyl-Ne-[N,α-(2- thiophenesuIfonyl)-L-phenyIalanyl]-L-lysine
The title compound was prepared from Nα-isobutyl-Nα-(4-nitrobenzenesulfonyl)-L-lysine hydrochloride (400 mg, 1.03 mmol, example 5, step C) as described in general procedure Be using Nα-(2-thiophenesulfonyl)-L-phenylalanine (311 mg, 1.2 mmol) which was prepared in step A of example 24. The final product was purified by preparative HPLC to yield 550 mg (80%) ofΛα-isobutyl-Λ7'α-(4-nitrobenzenesulfonyl)-Ne-[N'α-(2-thiophenesulfonyl)-L-phenylalanyl]-L- lysine. The latter derivative was hydrogenolysed following the indications of general procedure E. Purification by HPLC gave the desired material (400 mg, 65%).
Η ΝMR (DMSO-d6): δ 0J8 (q, J= 6.9, 6H), 1.00-1.11 (m, 4H), 1.32-1.35 (m, IH), 1.70-1.74 (m, IH), 1.83-1.88 (m, IH), 2.59-2.67 (m, 2H), 2.74-2.96 (m, 4H), 3.90 (t, J= 6.5, IH), 4.11 (t, J= 7.2, IH), 6.50 (d, J= 8.1, 2H), 6.95 (t, = 4.0, lH), 7.10-7.21 (m, 5H), 7.30 (d, = 4.0, lH), 7.34 (d, J= 8.1 , 2H), 7.77 (d, J= 4.0, IH), 7.81 (t, J= 5.3, IH), 8.22 (d, J= 8.1, IH). LC-MS: 649.8 (M - H)-, 98% pure.
Example 47. Preparation of Nα-(4-aminobenzenesuιfonyl)-Nα-isoburyl-Ne-[N'α-(4- methylbenzenesulfonyι)-L-asparagyl]-L-Iysine
Step A. Preparation Nα-(4~methylbenzenesulfonyl)-L-asparagine
L -asparagine was reacted with 4-methylbenzenesulfonyl chloride under the conditions used in general procedure A giving the title compound which was recrystallised from ether (40%>). 'H NMR (CDC13); δ 2.20-2.23 ( , IH), 2.33 (s, 3H), 2.42-2.50 (m, IH), 4.00-4.03 (br s, IH), 6.8 (s, IH), 7.26 (s, 2H), 7.51 (s, 2H).
Step B. • Preparation of Nα-(4-aminobenzenesulfonyI)-Nα-isobutyl-Ne-[N'α-(4- methylbenzenesulfonyl)-L-asparagyl]-L-lysine
The title compound was prepared from Nα-isobutyl-Nα-(4-nitrobenzenesulfonyl)-L-lysine hydrochloride (400 mg, 1.03 mmol, example 5, step C) as described in general procedure Be using Nα-(4-methylbenzenesulfonyl)-L-asparagine (286 mg, 1.0 mmol) which was prepared in step A this example. The final product, Nα-isobutyl-Nα-(4-nitrobenzenesulfonyl)-Ne-[N'α-(4- methylbenzenesulfonyl)-L-asparagyl]-L-lysine was subsequently hydrogenolysed following the indications of general procedure E. Purification by HPLC gave the desired material (185 mg, 76%).
Η ΝMR (CDC13): δ 0.78 (d, J= 6.3, 3H), 0.81 (d, J= 6.3, 3H), 0.94-1.13 (m, 4H), 1.32-1.35 (m, IH), 1.66-1.69 (m, IH), 1.83-1.88 (m, IH), 2.10-2.29 (m, 2H), 2.31 (s, 3H), 2.82-2.99 (m, 2H), 3.95 (t, J= 6.5, IH), 4.11 (t, J= 7.2, IH), 6.80 (d, J= 8.0, 2H), 7.19 (d, J= 7.9, 2H), 7.32 (d, J= 7.9. 2H), 7.64 (d, J= 8.0, 2H). LC-MS: 624.8 (M - H)\ 98% pure.
Example 48. Preparation of Λrα-isobutyI-Nα-(4-methyIbenzenesuIfonyl)-Ne-(N'α- benzyloxycarbonyI-L-asparagyI)-L-lysine
The title compound was prepared from solid phase bound Nα-isobutyl-Nα-(4- methylbenzenesulfonyl)-Λ?e-(9-fluorenylmethoxycarbonyl)-L-lysine as described in general procedure Bb using commercially available Nα-benzyloxycarbonyl-L-asparagine (319 mg, 1.2 mmol). The final product was purified by preparative HPLC to yield 46 mg (41%) of the desired material.
LC-MS: 603.1 (M - H)\ 98% pure. Example 49. Preparation of Nα-(4-aminobenzenesulfonyl)-Nα-isobutyl-Ne-[N'α-(2- thiophenesuIfonyI)-L-phenylaIanyl]-L-lysine hydrazide
A solution of Nα-(4-aminobenzenesulfonyl)-Nα-isobutyl-Ne-[N'α-(2-thiophenesulfonyl)-L- phenylalanyl]-L-lysine (650 mg, 1.0 mmol, example 46) dissolved in EtOAc (20 mL) was treated with />nitrophenol (139 mg, 1.0 mmol) and DCC (206 mg, 1.0 mmol) and was left in ' a refrigerator overnight. Afterwards, the precipitate was filtered off through celite and the solvent evaporated. The crude residue was used without further purification. A portion of the residue (55 mg, 0.072 mmol) was added to a solution of hydrazine hydrate in ethanol (IM, 10 mL). The resulting solution was stirred for 3 h before evaporation of the solvent. The residue was purified by preparative HPLC to yield 25 mg, 30% of the desired material.
LC-MS: 663.1 (M - H)", 95% pure.
Example 50. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-(Λr'α-benzoyl- L-phenyIaIanyl)-L-Iysine
Step A. Preparation of Nα-benzoyl -L-phenylalanine
L-phenylalanine was reacted with benzoyl chloride under the conditions used in general procedure A giving the title compound which was recrystallised from ether (60%).
'HΝMR (CDCL : δ 3.21-3.39 (m, 2H), 5.09 (q, = 6.6, IH), 7.10-7.27 (m, 5H), 7.42 (t, J= 6.9, 2H), 7.5 (t, J= 7.0, IH), 7.68 (d, J= 6.9, 2H). LC-MS: 268 (M - H)\ 99% pure.
Step B. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Λ/e-(N'α-benzoyl-L- phenylalanyl)-L-lysine
The title compound was prepared from solid phase bound Nα-isobutyl-Nα-(4- methylbenzenesulfonyl)-Ne-(9-fluorenylmethoxycarbonyl)-L-lysine as described in general procedure Bb using Nα-benzoyl-L-phenylalanine (321 mg, 1.2 mmol). The final product was purified by preparative HPLC to yield 44 mg (58%) of the desired material.
ΗNMR (CDC13): δ 0.83 (d, J= 6.9, 6H), 1.08-1.11 (m. 2H), 1.23-1.25 (m, 2H), 1.45-1.52 (m, IH), 1.89-1.99 (m, 5H), 2.31 (s, 3H), 2.94-3.19 (m, 6H), 4.24 (t, J= 6.9, IH), 4.89 (t, J= 5.9, IH), 7.19-7.26 (m, 7H), 7.30 (t, J= 7.9, 2H), 7.46 (t, J= 7.8, IH), 7.73-7.82 (m, 4H). LC-MS: 606.2 (M - H)\ 98% pure.
Example 51. Preparation of Nα-isoburyl-Nα-(4-methylbenzenesulfonyI)-Ne-[N'α-(4- morpholinecarbonyl)-L-phenylalanyl]-L-lysine
Step A. Preparation of Nα-(4-morpholinecarbonyl)-L-phenylalanine
L-phenylalanine was reacted with 4-morpholinecarbonyl chloride under the conditions used in general procedure A giving the title compound which was recrystallised from ether (69%>).
'H ΝMR (CDC13): δ 3.11-3.39 (m, 6H), 3.57 (s, 4H), 4,65 (q, J= 6.6, IH), 7.10-7.27 (m, 5H).
Step B. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-[N'α-(4- morpholinecarbonyl)-L-phenylalanyl]-L-lysine
The title compound was prepared from solid phase bound Nα-isobutyl-Λrα-(4- methylbenzenesulfonyl)-Ne-(9-fluorenylmethoxycarbonyl)-L-lysine as described in general procedure Bb usingNα-(4-morpholinecarbonyl)-L-phenylalanine (330 mg, 1.2 mmol). The final product was purified by preparative HPLC to yield 41 mg (53%) of the desired material.
LC-MS: 615.2 (M - H)\ 95% pure.
Example 52. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-(N'α-pivaloyl- L-phenylalanyl)-L-lysine
Step A. Preparation of Nα-pivaloyl-L-phenylalanine L-phenylalanine was reacted with pivaloyl chloride under the conditions used in general procedure A giving the title compound which was recrystallised from hexanes (40%>).
Η NMR (CDC13): δ 1.16 (s 9H), 3.12-3.31 (m, 2H), 4.85 (t, J= 5.1, IH), 7.11-7.32 (m, 5H). Step B. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-(N'α-pivaloyl-L- phenylalanyl)-L-lysine
The title compound was prepared from solid phase bound Nα-isobutyl-Nα-(4- methylbenzenesulfonyl)-Ne-(9-fluorenylmethoxycarbonyl)-L-lysine as described in general procedure Bb using Nα-pivaloyl-L-phenylalanine (300 mg, 1.2 mmol). The final product was purified by preparative HPLC to yield 49 mg (66%) of the desired material.
LC-MS: 586.1 (M - H)\ 95% pure.
Example 53. Preparation of Λrα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-[N'α-(2- thiophenesulfonyl)-L-tryptophanyl]-L-Iysine
Step A. Preparation of Nα-(2-thiophenesulfonyl)-L-tryptophan
L-tryptophan was reacted with 2 -thiophenesulfonyl chloride under the conditions used in general procedure A giving the title compound which was recrystallised from DCM (32%).
Η ΝMR (CDCI3): δ 2.9-3.1 1 (m, 2H), 3.91 ( ,y= 7.0, IH), 6.86-7.01 (m, 4H), 7.21-7.3 (m, 3H), 7.70 (s. IH). 8.38 (s. IH).
Step B. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-[N'α-(2- thiophenesulfonyl)-L-tryptophanyI]-L-lysine
The title compound was prepared fromNα-isobutyl-N -(4-methylbenzenesulfonyl)-L-lysine (100 mg. 0.29 mmol, example 1. step E) as described in general procedure Be using Nα-(2- thiophenesulfonyl)-L-tryptophan (90 mg, 0.3 mmol) which was prepared in step A of this example. The final product was triturated with ether to yield 12 mg (6%>) of the desired material.
LC-MS: 687.2 (M - H)\ 85% pure.
Example 54. Preparation of Nα-isobutyI-Nα-(2-thiophenesuIfonyI)-Ne-[N,α-(4- methylbenzenesuIfonyI)-L-phenylalanyl]-L-Iysine
Step A. Preparation of Nα-isobutyI-Nα-(2-thiophenesulfonyl)-L-α-amino-e-caprolactam
Nα-isobutyl-L-α-amino-e-caprolactam (example 1, step C) (2.56 g, 14.0 mmol, free base) was dissolved in DCM (50 mL) and treated with diisopropylethylamine (4.0 mL, 20 mmol) followed by freshly recrystaUized 2-thiophenenesulfonyl chloride (2.56 g, 14.0 mmol). The mixture was stirred overnight (TLC shows the reaction to be complete after 2 h). The solution was extracted with IN HCl and the organic layer was dried and evaporated. Then, the residue was dissolved in boiling MeOH (150 mL) and placed in the refrigerator for 3 h. The thin needles obtained were filtered off and air dried giving 3.6 g (78%) of pure product. This compound was used without further purification in the next step.
Step B. Preparation of Nα-isobutyl-Nα-(2-thiophenesulfonyl)-L-lysine hydrochloride
A mixture of Λfa-isobutyl-Λtø-(2-thiophenesulfonyl)-L-α-amino-e-caprolactam (3.5 g, 10.1 mmol), AcOH (12 mL) and 6Ν HCl (50 mL) was refluxed for 6 h until all solids had disappeared. Afterwards, the solution was evaporated to give 2.8 g, 71% of the hydrochloride salt.
•H NMR (DMSO-d6): δ 0.72 (dd, J= 5.8, 6.4, 6H), 1.13-1.17 (m, 2H), 1.42-1.46 (m 2H), 1.79- 1.87 (m, 2H), 2.67 (t, J= 7.2, 2H), 2.80-2.91 (m, 2H), 4.13 (t,J= 7.2, IH), 7.11 (t, .7= 5.1, IH), 7.50 (d, J= 5.5, IH), 7.85 (d, J= 5.6, IH).
Step C. Preparation of Nα-isobutyl-Nα-(2-thiophenesulfonyl)-Ne-[N 'α-(4- methylbenzenesulfonyl)-L-phenylalanyl]-L-lysine A suspension of Nα-isobutyl-Nα-(2-thiophenesulfonyl)-L-lysine hydrochloride (150 mg, 0.5 mmol), in THF (20 mL) was treated with a IN ΝaOH (0.5 mL) to pH 10. A solution of commercially available Nα-(4-methylbenzenesulfonyl)-L-phenylalanine'acid chloride (150 mg, 0.4 mmol), in dry THF (10 mL) was added to the suspension and stirred for 4 h. Afterwards, water (2 mL) was added resulting in a clear solution. Then, EtOAc (30 mL) was added and the organic phase was washed with IN HCl. The organic phase was removed. Evaporation of the solvent gave a crude product which was purified by preparative HPLC to yield 150 mg (57%) of the title compound.
ΗΝMR (DMSO-d6): δ 0.79 (d, 7= 6.3, 3H), 0.82 (d, 7= 6.3, 3H), 1.00-1.1 1 (m, 4H), 1.23-1.25 (m, IH), 1.80-1.84 (m, IH), 1.89-1.98 (m, IH), 2.32 (s, 3H), 2.59-2.67 (m, 2H), 2.84 and 2.95 (ABX, 7= 15.5, 4.8, 2H)„3.88 (t, 7= 6.0, IH), 4.21 (t, 7= 6.9, IH), 7.04 (d, 7= 7.6, 2H), 7.12- 7.21 (m, 5H), 7.35-7.44 (m, 3H), 7.87 (d, 7= 7.61, 2H). LC-MS: 648.5 (M - H)\ 98% pure.
Example 55. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-[N'α-isobutyl- Λ7'α-(4-methyIbenzenesulfonyl)-glycyl]-L-lysine
Step A. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-glycine
The title compound was prepared in a three-step sequence from rert-butyl bromoacetate. Initially, tert-butyl bromoacetate (1.91 g, 10 mmol) dissolved in isobutylamine (60 mmol) was stirred at room temperature for 2 h. The reaction mixture was filtered and the excess isobutylamine was distilled yielding 80%> of pure N-isobutyl gl cine tert-butyl ester. Secondly, the intermediate was reacted with 4-methylbenzenesulfonyl chloride (8 mmol) as described for the preparation of Na- isobutyl-Nα-(4-methylbenzenesulfonyl)-L-α-amino-e-caprolactam presented in example 1 (step D). In this case, triethylamine was used instead of diisopropylethylamine . Thirdly, deprotection of the tert-butyl ester with TFA provided the final product quantitatively (two last steps).
'H ΝMR (CDC13): δ 0.80 (br s, 6H), 1.75-1.82 (m, IH), 2.32 (s, 3H), 2.85-2.9 (br s, 2H), 3.83 (s, 2H), 7.26 (d, 7= 7.9, 2H), 7.6 (d, 7= 7.9..2H). LC-MS: 286 (M - H)\ 99% pure. Step B. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-[N'α-isobutyl-N'α-(4- methylbenzenesulfonyl)-glycyl]-L-lysine
The title compound was prepared from Nα-isobutyl-Nα-(4-methy lbenzenesulfonyl)-L-lysine (100 mg, 0.29 mmol, example 1 , step E) as described in general procedure Be using Nα-isobutyl-Nα- (4-methylbenzenesulfonyl)-glycine (90 mg, 0.3 mmol) which was prepared in step A of this example. The final product was triturated with ether to yield 40 mg (21%) of the desired material.
LC-MS: 623.8 (M - Hj, 95% pure.
Example 56. Preparation of Nα-(4-aminobenzenesulfonyl)-Nα-isobutyl-Ne-JN,α-(4- acetylaminobenzenesulfonyl)-L-phenylaIanyl]-L-Iysine
The title compound was prepared from Nα-(4-aminobenzenesulfonyl)-Nα-isobutyI-L-Iysine (100 mg, 0.29 mmol, example 166, step B) as described in general procedure Be using Nα-(4- acetamidobenzenesulfonyl)-L-phenylalanine (110 mg, 0.3 mmol) which was prepared in step A of example 3. The final product was purified by HPLC to give 50 mg (23%) of pure adduct.
LC-MS: 700.8 (M - H)\ 95% pure.
Example 57, Preparation of Nα-isobutyI-Nα-(4-methyIbenzenesulfonyl)-Ne-[N,α-(2- thiophenesuIfonyl)-L-phenylalanyl]-L-lysine methyl ester
The title compound was prepared by treating Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne- [N'α-(2-thiophenesulfonyl)-L-phenylalanyl]-L-lysine (60 mg, 0.1 mmol, example 24) dissolved in MeOH (2 mL) with DCC (1 eq.). The reaction mixture was stirred at room temperature for a period of 2 h. Filtration and evaporation of the solvent followed by HPLC purification gave the desired methyl ester (15 mg, 22%).
LC-MS: 662.1 (M - H)", 99% pure. Example 58. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-[N'α-(2- thiophenesulfonyl)-L-phenylalanyl]-L-lysinamide
A solution of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-[N'α-(2-thiophenesulfonyl)-L- phenylalanyl]-L-lysine (650 mg, 1.0 mmol, example 24) dissolved in EtOAc (10 mL) was treated with / nitrophenol (139 mg, 1.0 mmol) and DCC (206 mg, 1.0 mmol) and Was left in 'a refrigerator overnight. Afterwards, the precipitate was filtered off through celite and the solvent evaporated. The crude intermediate was used without further purification in the next step. A portion of the intermediate (25 mg, 0.03 mmol) was added to a solution of ammonia in ethanol (IM, 10 mL). The resulting solution was stirred for 3 h before evaporation of the solvent. The residue was purified by preparative HPLC to yield 2.6 mg, 11% of the desired material.
LC-MS: 647.2 (M - H)\ 99% pure.
Example 59. Preparation of Nα-isoburyl-Nα-(4-mcthylbenzenesulfonyl)-Ne-[N'α-(2- thiophenesulfonyl)-L-phenylalanyl]-L-lysineN-hydroxylamide
A portion of the crude intermediate of example 58 above, (25 mg, 0.03 mmol) was added to a solution of hydroxy] amine in ethanol (IM, 10 mL). The resulting solution was stirred for 3 h before evaporation of the solvent. The residue was purified by preparative HPLC to yield 4.0 mg, 18% of the desired material.
LC-MS: 663.1 (M - H)\ 99% pure.
Example 60. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-[N'α-(2- thi oph en esulf onyl)-L-ph enylalanyl] -L-lysin e ethanolamide
A portion of the crude intermediate of example 58 above, (25 mg, 0.03 mmol) was added to a solution of ethanolamine in ethanol (IM, 10 mL). The resulting solution was stirred for 3 h before evaporation of the solvent. The residue was purified by preparative HPLC to yield 15 mg, 21% of the desired material. LC-MS: 691.2 (M - H)\ 99% pure.
Example 61. Preparation of Nα-isoburyl-Nα-(4-methyIbenzenesulfonyl)-Ne-[N'α-(4- methylbenzenesulfonyl)-L-asparagyl]-L-lysine
The title compound was prepared fromNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine (100 mg, 0.29 mmol, example 1, step E) as described in general procedure Be using Nα-(4- methylbenzenesulfonyl)-L-asparagine (90 mg, 0.3 mmol) which was prepared in step A of example 47. The final product was triturated with ether to yield 95 mg (15%) of the desired material.
]H ΝMR (CDC13): δ 0.78 (d, 7= 6.3, 3H), 0.81 (d, 7= 6.3, 3H), 0.99-1.03 (m, 4H), 1.32-1.35 (m. lH), 1.66-1.69 (m, lH), 1.83-1.88 (m, lH), 2.10-2.29 (m, 2H), 2.35 (s, 3H), 2.38 (s, 3H), 2.82 and 2.99 (m, 2H), 3.95 (t, 7= 6.5, IH), 4.11 (t, 7= 7.2, IH), 7.19 (d, 7= 7.9, 2H), 7.32 (d, 7= 7.9, 2H), 7.55-7.64 (m, 4H). LC-MS: 625.8 (M + H)\ 97% pure.
Example 62. Preparation of Nα-isobutyl-Λrα-(4-methyIbenzenesuIfonyl)-Ne-(N'α-pivaloyl- L-asparagyI)-L-lysine
Step A. Preparation of Nα-pivaloyl-L-asparagine
L-asparagine was reacted with pivaloyl chloride under the conditions used in general procedure A giving the title compound which was recrystallised neat (80%). This compound was used without further purification in the next step.
Step B. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-(N'α-pivaloyl-L- asparagyl)-L-lysine
The title compound was prepared from Nα-isobutyI-Nα-(4-methyIbenzenesulfonyl)-L-ly sine (100 mg, 0.29 mmol, example 1 , step E) as described in general procedure Be using Nα-pivaloyl-L- asparagine (65 mg, 0.3 mmol) which was prepared in step A this example. The final product was purified by preparative HPLC to yield 21 mg (12%) of the desired material.
LC-MS: 555.7 (M + H)+, 90% pure. \ '
Example 63. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-(N,α-benzoyl- L-asparagyI)-L-lysine
Step A. Preparation of Nα-benzoyl-L-asparagine ' .
L-asparagine Was reacted with benzoyl chloride under the conditions used in general procedure A giving the title compound which was recrystallised neat (90%). This compound was used without further purification in the next step. •
Step B. Preparation of Λ7α-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-(N'α-benzoyl-L- ' asparagyl)-L-Iysine ,
The title compound was prepared from Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine (100 mg, 0.29 mmol, example 1, step E) as described in general procedure Be using Nα-benzoyl-L- asparagine (70 mg, 0.3 mmol) which was prepared in step A this example. The final product was purified by preparative HPLC to yield 14 mg (9%>) of the desired material.
LC-MS: 575.2 (M + H)*, 99% pure. '
Example 64. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-[N'α-(2- thiophenesulfonyl)-L-phenylalanyl]-L-lysme hydrazide
A portion of the crude intermediate of example 58 above, (50 mg, 0.06 mmol) was added to a solution of hydrazine in ethanol (IM, 10 mL). The resulting solution was stirred for 3 h before' evaporation of the solvent. The residue was purified by preparative HPLC to yield 25 mg, 60% of the desired material. LC-MS: 664.2 (M - H)\ 99% pure. ^ :
Example 65. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-[l,2,3,4- tetrahydroisoquinoline-N'-(4-methylbenzenesulfonyl)-3-carbonyl]-L-lysine
This particular preparation is based in scheme 4 of this invention.
Step A. Preparation of Nα-isobύtyl-Ne-benzyloxycarbonyl-L-lysine methyl ester
To a stirred solution of commercially available Ne-benzyloxycarbonyl-L-lysine methyl ester hydrochloride (9.92 g, 30 mmol), AcOH (6 mL) and ΝaCΝBH3 (33 mmol) in MeOH (250 mL) at 0°C was added a solution of isobutyraldehyde (3.01 mL, 33 mmol) in MeOH (80 mL); The solution was warmed to room temperature and stirred for 2 h. A saturated solution of K2CO3 (150 mL) was added and the solution was decanted from the solid and coevaporated on vacuo. The residue was partitioned between EtOAc (300 mL) and H2O (200 L). The organic layer was washed with K2CO3 (IM) and with brine, then dried and concentrated. The crude was used in the next step without further purification.
J : : ' .
1 ! !
Step B. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-benzyloxycarbonyl-L- lysine methyl ester ; ' ■ i . ■ .
To a stirred solution of Nα-isobutyl-Ne-benzyloxycarbonyl-L-lysine methyl ester (336 mg, 1 mmol) in CH2C12 (2 mL) was added 4-methylbenzenesulfonyl chloride (286 mg, 1.5 mmol) and triethylamine (174 μL, 1' mmol). The reaction mixture was allowed to stir for 3 days, then it was diluted with IN HCl and extracted with CH2C12. The organic layer was dried (MgSO4) and concentrated. The crude was flash chromatographed using hexane EtOAc as eluent to obtain the corresponding sulfonamide. '
Yield: 71% (steps A and B) Η NMR (DMSO-d6): δ 0.84 (d, 7= 7.2, 3H), 0.86 (d, 7= 6.3, 3H), 1.30-1.68 (m, 5H), 1.88-2.00 (m, 2H), 2.42 (s, 3H), 2.92 and 3.00 (ABX, 7= 14.7, 8.2, 2H), 3.18 ( , 2H), 3.50 (s, 3H), 4.40 (t, 7= 7.4, IH), 4J8 (br s, IH), 5.11 (s, 2H), 7.27-7.71 (m, 9H).
Step C. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine methyl ester
Pd/C 10% (120 mg) was added to a solution of the above sulfonamide (491 mg, 1 mmol) in EtOAc/MeOH (3 mL/ 3 mL). The suspension was flushed with H2 and maintained under H2 pressure until complete consumption of the starting material. The insoluble material was filtered off, and the filtrate was concentrated under reduced pressure to give the desired amine in quantitative yield. This compound was used without purification in the next step.
Figure imgf000092_0001
added ΝaOH (0.6 mmol). The reaction mixture was stirred until complete consumption of the starting ester, then diluted with IN HCl until acidic pH and extracted with EtOAc. The organic phase was dried (MgSO4) and concentrated to give the desired acid in quantitative yield.
Overall yield: 62% (steps C, D and E)
Η NMR (DMSO-d6): δ 0.80 (d, 7= 6.5, 3H), 0.83 (d, 7= 6.6, 3H), 1.00-1.30 (m, 4H), 1.35 (s, 9H), 1.42'(m, IH), i .70-1.92 (m, 2H), 2.35 (s, 3H), 2.80-3.10 (m, 6H), 4.12 (m, IH), 4.35 (m,
Figure imgf000093_0001
'HΝMR (DMSO-d6): δ 0.80 (d,7= 6.5, 3H), 0.83 (d,7= 6.8, 3H), 1.22-1.57 (m, 5H), 1.82-1.98 (m, 2H), 2.38 (s, 3H), 2.88-3.22 (m, 5H), 3.30 (d, 7= 16.5, IH), 4.10-4.40 (m, 4H), 7.25 (m, 4H), 7.40 (d, 7= 7.4, 2H), 7.69 (d, 7= 7.5, 2H), 8.60 (s, IH), 9.40 (br s, IH), 9.63 (br s, IH).
' i ' I
•i : i ' ' '
Step G. Preparation of Nα-isobutyl-Nα-(4-methy lbenzenesulfonyl)-Ne-[l, 2,3,4- i ■ • ' < • tetrahydroisoquinoline-N,-(4-methylbenzenesulfonyl)-3-carbonyl]-L-lysine x ■ • .
The final product was prepared from Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-[l, 2,3,4- tetrahydroisoquinoline-3 -carbonyl]-L- lysine trifluoroacetic acid salt (step F above) following the indications of step B of this example. The desired material was obtained in 71%) yield.'
ΗΝMR (DMSO-d6): δ 0.80 (d, 7= 6.5, 3H), 0.83 (d, 7= 6.8, 3H), 1.00-1.40 (m, 5H), 1.68-1.90 (m, 2H), 2.33 (s, 3H), 2.35 (s, 3H), 2.70-3.00 (m, 6H), 4.12 (m, IH), 4.42-4.53 (m, 3H), 7.10-7.40 (m, 8H), 7.63 (d, 7 = 8.2, 4H), 7.90 (m, IH), 12.70 (br s, IH).
Example 66. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-[N,α-(4- methylbenzenesulfonyl)-D-phenylalanyl]-D-lysine
The title compound was prepared in the same manner as in the previous example (example 65) using Nα-isobutyl-Ne-benzyloxycarbonyl-D-lysine methyl ester and 4-methylbenzenesulfonyl-D- phenylalanine as the starting materials.
Figure imgf000094_0001
The title compound' was prepared in the same manner as in the example 65 using Nα-isobutyl-Ne- benzyloxycarbonyl-D-lysine methyl ester and 4-methylbenzenesulfonyl-L-phenylalanine as the starting materials. '
'HMMR CCDCy: δ 0.77 (d, 7= 6.3, 3H), 0.80 (d, 7= 6.3, 3H), 1.00-1.11 (m, 4H), 1.23-1.25 (m, IH), 1.70-1.74 (m, IH), 1.89-1.93 (m, IH), 2.30 (s, 3H), 2.32 (s, 3H), 2.59-2.67 (m, 2H), 2.87 and 2.93 (ABX, 7= 14.1!, 4.2, 2H), 3.85 (t, 7= 5.9, IH), 3.63 (t, 7= 6.9, IH), 6.90-7.10 (m, 7H), 7.24 (d, 7= 8.6, 2H), 7.44 (d, 7= 8.1, 2H), 7J3 (d, 7= 8.1, 2H). : Example 68. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-[N'α-(4- methylbenzenesulfonyl)-L-phenylalanyl]-L-lysine methyl ester
To a stirred solution'of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine methyl ester (369 mg, lmmol, example 65, step C) in THF K2CO3(lM) (3 mL/3 mL) was added Nα-(4- methylbenzenesulfonyl)-L-phenylalanine N-hydroxysuccinimide ester (500 mg, 1.2 mmol). The reaction mixture was stirred overnight, then diluted with IN HCl and extracted with EtOAc. The organic layer was dried (MgSO4) and concentrated. The crude was purified by flash chromatography using hexane/EtOAc as eluent to afford the desired product (77% yield).
ΗΝMR (DMSO-d6): δ 0.83 (d, 7= 7.0, 3H), 0.86 (d, 7= 6.8, 3H), 1.22-1.50 (m, 4H), 1.60 (m, IH), 1.80-1.95 ( , 2H), 2.40 (s, 3H), 2.42 (s, 3H), 2.85-3X5 ( , 4H), 3.12 ( , 2H), 3.50 (s, 3H), 3.88 (m, 3H), 3.49 (t, 7=:5.0, IH), 5.22 (m, 1H),,6.42 (t,7= 5.0, IH), 6.96 (d,7= 8.0, 2H), 7.12- 7.20 (m, 5H), 7.30 (d, 7= 8.0, 2H), 7.51 (d, 7= 7.5, 2H), 7.72 (d, 7= 7.8, 2H).
Example 69. Preparation of Nα-isoburj'l-7Vα-(4-methylbenzenesulfonyl)-Ne-[N'α-(4- methyIbenzenesulfonyl)-L-tryptophanyl]-L-lysine methyl ester
To a stirred solution of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine methyl ester (369- mg, lmmol, example 65, step C) in THF/K2CO3 (IM) (3 mL/3 mL) was added Nα-(4- methylbenzenesulfonyl)-L-tιyptophan N-hydroxysuccinimide ester (547 mg, 1.2 mmol). The reaction mixture was stirred overnight, then diluted with IN' HCl and extracted with EtOAc. The organic layer was dried (MgSO4) and concentrated. The crude was purified by flash chromatography using hexane/EtOAc as eluent to afford the desired product (71% yield).
Η NMR (DMSO-d6): δ OJ7 (t, 7= 7.5, 6H), 1.00-1.10 (m,4H), 1.40 (m, IH), 1.70 (m, IH), 1.85 (m, IH), 2.29 (s, 3H), 2.38 (s, 3H), 2.87-3.00 (m, 3H), 3.44 (s, 3H), 3.85 (m, IH), 4.24 (t, 7= 7.3, IH), 5.74 (s, 2H), 6.88-7.10 (m, 3H), 7.15 (d, 7= 8.2, 2H), 7.25-7.46 ( , 6H), 7.65 (d, 7= 8.2, 2H), 7.75 (br s, IH), 7.84 (d, 7= 8.6, IH), 10.71. (s, IH). Example 70. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-(N'α-benzoyl- L-phenylalanyl)-L-lysme methyl ester
To a stirred solution of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine methyl ester (369 mg, lmmol, example 65, step C) in THF/K2CO3(lM) (3 mL/3 mL) was added Nα-benzoyl-L- phenylalanine N-hydroxysuccinimide ester (440 mg, 1.2 mmol). The reaction mixture was stirred overnight, then diluted with IN HCl and extracted with EtOAc. The organic layer was dried (MgSO4) and concentrated. The crude was purified by flash chromatography using hexane/EtOAc as eluent to afford the desired product (82% yield).
ΗΝMR (DMSO-d6): δ 0.83 (d,7= 6.3, 3H), 0.85 (d,7= 6.8, 3H), 1.22-1.50 (m, 4H), 1.55-1 JO (m, IH), 1.80-2.00 (m, 2H), 2.43 (s, 3H), 2.90 and 3.00 (ABX, 7= 14.0, 7.5, 2H), 3.12-3.30 (m, 4H), 3.50 (s, 3H), 4.39 (m, IH), 4.82 (m, IH), 5.95 (br s, IH), 6.98 (br s, IH), 7.20-7.52 (m, 1 OH), 7.65-7.75 (m, 4H).
! '■■ ' I : " ' •
Example 71. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyI)-Ne-[N,α-(4- methylbenzenesulfonyl)-L-thiophenylalanyl]-L-lysme methyl ester
The title compound was prepared fromNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-[N'α-(4- ' ■ ! i < methylbenzenesulfony l)-L-phenylalanyl]-L-lysine methyl ester, product of example 68, following the indications of general procedure F. The thioamide was Obtained in 64%) yield.
!H MR (DMSO-d6): 6 0.83 (d, 7= 6.8, 3H), 0.86 (d, 7= 6.8, 3H), 1.30 (m, 2H), 1.48-1.60 (m, 2H), 1.70 (m, IH), 1.80-1.95 (m, 2H), 2.42 (s, 3H), 2.46 (s, 3H), 2.88 and 3.20 (ABX, 7= 13.5, 7.5, 2H), 3.00 (m, 2H), 3.45 (m, 2H), 3.52 (s, 3H), 4.18 (m, IH), 4.40 (m, IH), 5.37 (d, 7= 7.0, IH), 7.00 (d, 7= 8.0, 2H), 7.18 (m, 5H), 7.32 (d, 7= 7.5, 2H), 7.50 (d, 7= 7.5, 2H), 7.75 (d, 7= 8.0, 2H), 7.97 (br s, IH). , Example 72. Preparation of Nα-isobutyI-Nα-(4-methylbenzenesulfonyl)-Ne-[N,α-(4- methylbenzenesulfonyl)-L-thiophenylalanyl]-L-lysine
Saponification of ; Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-[N'α-(4- methylbenzenesulfony l)-L-thiophenylalanyl]-L-lysine methyl ester (example 71) using the conditions described in example 65 (step E) yielded 87% of the desired material.
Η ΝMR (DMSO-d6): δ 0.80 (d, 7= 6.5, 3H), 0.83 (d, 7= 6.8, 3H), 1.00-1.42 (m, 5H), 1.78 (m, IH), 1.90 (m, IH), 2.33 (s, 3H), 2.37 (s, 3H), 2.75 (m, IH), 2.82-3.15 9m, 6H), 4.15 (t, 7= 7:2, IH), 4.30 (m, IH), 7.00-7.25 (m, 7H), 7.38 (d,7= 8.2, 2H), 7.50 (d,7= 8.3, 2H), 7.68 (d,7= 8.2, 2H), 7.75 (d, 7= 9.0, IH), 9J7 (s, IH), 12.70 (br s, IH).
Example 73. Preparation of Nα-isobutyI-Nα-(4-methylbenzenesuIfonyl)-Ne-[N'α-(4- methylbenzenesulfonyl)-L-thiotryptophanyl]-L-lysine
Saponification of ; Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne- [N 'α-(4-
X i ! ■ : . ! ' methylbenzenesulfonyl)-L-thiotryptophanyl]-L-lysine ' methyl ester (example 75) using the conditions described in example 65 (step E) yielded 88% of the desired material.
,HΝMR (DMSO-d6): δ 0.80 (d, 7= 6.5, 3H), 0.83 (d, 7= 6.8, 3H), 1.00-1.42 (m, 5H), 1.73 (m, IH), 1.83 (m, IH), 2.28 (s, 3H), 2.37 (s, 3H), 2.80-3.02 (m, 3H), 3.03-3.15 (m, 3H), 4.15 (t, 7= 6.5,' IH), 4.30 (m, IH), 6.90 (t, 7= 7.4, IH), 7.00 (t, 7= 7.4, IH), 7.10 (s, IH), 7.20 (d, 7= 8.0, 2H), 7.28 (d,7= 8.2, IH), 7.30-7.42 (m, 4H), 7.67 (m, 4H), 9.68 (s, IH), 10.72 (s, lH), 12.76 (br s, lH). ' ; ;
Example 74. Preparation of Nα-isobutyl-Nα-(4-methyIbenzenesuIfonyI)-Ne-(N'α- benzenesulfonyl-L-thiotryptophanyl)-L-lysine
The title compound was prepared from Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-(N'α- benzenesulfonyl-L-tryptophanyl)-L-lysine, product of example 4, following the indications of general procedure F. The thioamide was obtained in 34% yield. Η NMR (DMSO-d6); δ 0.80 (d, 7= 6.5, 3H), 0.83 (d, 7= 6.8, 3H), 1.00-1.42 (m, 5H), 1.73 (m, IH), 1.83 (m, IH), 237 (s, 3H), 2.85-3.02 (m, 3H), 3.03-3.15 (m, 3H), 4.15 (t,7=,6.5, 1H), 4.30 (m, IH), 6.90 (t, 7= 7.4, IH), 7.00 (t, 7= 7.4, IH), 7.10 (s, IH), 7.27 (d, 7= 8.0, 2H), 7.32-7.50 (m, 5H), 7.55 (d, 7= 8.5, 2H), 7.67 (d, 7= 8.5, 2H), 7.81 (d, 7= 8.5, 2H ), 9.69 (s, IH), 10.72 (s, IH), 12.76 (br s, IH). ,
Example 75. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-[N,α-(4- methylbenzenesulfonyl)-L-thiotryptophanyl]-L-lysine methyl ester
The title compound was prepared fromNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-[N'α-(4- methylbenzenesulfony l)-L-tryptophanyl] -L-lysine methyl ester, product of example 69, following the indications of general procedure F. The thioamide was obtained in 55% yield.-
Η ΝMR (CDCl3): δ 0.83 (d, 7= 6.8, 3H), 0.87 (d, 7= 6.8, 3H), 1.20-1.62 (m, 5H), 1.80-1.98 (m, 2H), 2.33 (s, 3H), 2.44 (s, 3H), 2.88 and 3.02 (ABX, 7= 13.5, 7.5, 2H), 3.22 and 3.33 (ABX, 7 = 14.0, 7.5, 2H), 3.40-3.52 (m, 2H), 3.49 (s, 3H), 4.25 (m, IH), 4.39 (m, IH), 5.30 (s, IH), 5.40 (d, 7= 6.0, IH), 7.00-7.20 (m, 5H), 7.25-7.50 (m, 5H), 5J0 (d, 7= 7.5, 2H), 7.90 (t, 7= 5.0, l.H), 8.48 (s, lH). ' ' '' 1 ' ' ' ,
Example 76. Preparation of Nα-isobutyl-Nα-(47methylbenzenesulfonyl)-Ne-(N,α- thiobenzoyl-L-thiophenylalanyl)-L-lysine methyl ester
The title compound was prepared from Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-(N'α- benzoyl-L-phenylalanyi)-L-ly sine methyl ester, product of example 70, following the indications of general procedure F. The thioamide was obtained in 81 %> yield.
Η MR (DMSO-d6): δ 0.83 (d,7= 6.3, 3H), 0.86 (d,7= 6.8, 3H), 1.12-1.50 (m, 4H), 1:55-1.70 (m, IH), 1.80-2.00 (m, 2H), 2.45 (s, 3H), 2.87 (dd, 7= 14.0, 7.5, 2H), 2.98-3.10 (m, 2H), 3.40- 3.50 (m, 2H), 3.51 (s, 3H), 3.70 (m, IH), 4.39 (m, IH), 5.50 (br s, IH), 7.20-7.50 (m, 10H), 7.65- 7.75 (m, 4H), 9.15 (br s, IH). Example 77. Preparation of Nα-isobutyl-Nα-^-methylbenzenesulfonyrj-Ne^N'α- thiobenzoyl-L-thiophenylalanyl)-L-lysine
Saponification of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-(N'α-thiobenzoyl-L- thiophenylalanyl)-L-lysine methyl ester (example 76) using the conditions described in example 65 (step E) yielded 80% of the desired material.
• !HΝMR(DMSO-d6): δ 0.81 (t,7= 6.5, 6H), 1.15-1.52 (m, 5H), 1.80-1.90 (m, 2H), 2.36 (s, 3H), 2.92 and 2.98 (ABX, 7=' 14.5, 7.5, 2H), 3.10 (m, IH), 3.30-3.50 (m, 3H), 4.20 (t, 7= 7.0, IH), 5.60 (m, IH), 7.18-7.28 (m, 3H), 7.32-7.50 (m, 5H), 7.68 (m, 4H), 10.23 (m, 2H), 12.80 (br s,
IH). :, • ; : • :
Example 78. Preparation of Nα-isobutyI-Nα-(4-methylbenzenesulfonyI)-Ne-(N,α-tert- butoxycarbonyl-L-phenylalanyl)-L-lysine
The title compound was prepared from Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine hydrochloride (1.2 g, 3.1 mmol, example 1,' step E) suspended in THF (50 mL) and lΝ ΝaOH (0.5 mL. to reach pH 10) as described in general procedure Be using commercially available Nα- . benzyloxycarbonyl-L-phenylalanine (1.0 g, 3.5 mmol), N-hydroxysuccinimide (0.4 g, 3.5 mmol) and DCC (1.1 g, 4.8 mmol). The final product was triturated with ether to yield 1.61 g (95%) of the desired material.
ΗΝMR(CDC13): δ 0.83 (m, 6H), 1.08-1.11 (m, 2H), 1.23-1.25 (m,2H), 1.35 (s, 9H), 1.45-1. "52 (m, IH), 1.89-1.99 (m, 2H), 2.36 (s, 3H), 2.94-3.09 (m, 6H), 4.20-4.23 (m, 2H), 7.09-7.26 (m, 7H), 7.73 (d, 7= 8.1', 2H). LC-MS: 602.2 (M - H)", 95% pure.
Example 79. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-(L- phenylalanyl)-L-lysine >
This product was prepared from Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-(N'α-tert- butoxycarbonyl-L-phenylalanyl)-L-lysine (600 mg, 1.0 mmol, example 78) as described in general procedure C. The residue was triturated with ether and placed under high vacuum to yield a hard white foam (440 mg, 88%). ' '
'H NMR (CDC13): δ 0J8 (d, 7= 6.3, 3H), 0.81 (d, 7= 6.3, 3H), 1.00-1.11 ( , 2H), 1.23-1.25 (m, 2H), 1.44-1.47 (m, IH), 1J0-1J4 (m, IH), 1.89-1.93 (m, IH), 2.32 (s, 3H), 2.82-2.89 (m, 2H), 2.91-2.95 (m, 2H), 3.88 (br s, IH), 4.17 (t, 7= 5.9, IH), 7.14 (d, 7= 8.0, 2H), 7.23 (t, 7= 4.9, IH), 7.29 (t, 7= 4.8, 2H), 7.31 (d, 7= 5.0, 2H), 7.64 (d, 7= 8.1, 2H), 8.05 (br s, 3H). LC- MS: 502.2 (M - H)', 95% pure. "
Example 80. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-[N,α- (carbotetrahydro-3-furanyloxy)-L-phenylalanyl]-L-lysine
Step A. Preparation of Nα-(carbotetrahydro-3-furanyloxy)-L-phenylalanine
' <
L-phenylalanine was reacted with tetrahydro-3-furanyloxy-l-nitroρhenyl carbonate under the conditions used in general procedure A giving the title compound which was recrystallised from DCM (28%). This compound was used without further purification in the next step.-
Ste B. Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-[N'α-(carbotetrahydro-3- f ranyloxy)-L-phenylalanyI]-L-lysine '•
The title compound was prepared fromNα-isobufyl-Nα-(4-methylbenzenesulfonyl)-L-lysine (100 mg, 0.29 mmol, example 1, step E) as described in general procedure Be using Nα- (carbotetrahydro-3-fliranyloxy)-L-phenylalanine (84 mg, 0.3 mmol) which was prepared in step A of this example. The final product was triturated with ether to yield 51 mg (27%>) of the desired material.
LC-MS: 618.9 (M + H)+, 90% pure'. Example 81. Preparation of Nα-isobutyl-Nα-(4-methyIbenzenesuIfonyI)-Ne-(N'α-ϊert- butoxyearbonyl-N'α-methyl-L-phenylalany^-L-Iysine
The title compound was prepared fromNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine (100 mg, 0.29 mmol, example 1, step E) as described in general procedure Be using commercially available Nα-tert-butoxycarbonyl-Nα-methyl-L-phenylalanine (83 mg, 0.3 mmol). The final product was triturated with ether to yield 176 mg (90%>) of the desired material.
LC-MS: 616.8 (M - H)γ94% pure.
Example 82. Preparation of Nα-isoburyl-Nα-(4-methylbenzenesulfonyl)-Ne-(N,α-/'er - butoxycar,bonyl-L-methionyl)-L-lysine
The title compound was prepared from Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine (100 mg, 0.29 mmol, example 1, step E) as described in general procedure Be using commercially available Nα-tert-butoxycarbonyl-L-methionine (75 mg, 0.3 mmol). The final product was triturated with ether to yield 170 mg (96%) of the desired material.
LC-MS: 586.8 (M - H)\ 90% pure.
Example 83. Preparation of Nα-isobutyI-Nα-(4-methyIbenzenesulfonyI)-Ne-[N'α-tert- butoxycarbonyl-S-(4-methylbenzyl)-L-cysteinyl]-L-lysine
The title compound was prepared fromNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine (100 mg, 0.29 mmol, example 1, step E) as described in general procedure Be using commercially available Nα-tert-butoxycarbonyl-S-(4-methylbenzyl)-L-cysteine (67 mg, 0.3 mmol). The final product was triturated with ether to yield 130 mg (65%) of the desired material.
LC-MS: 562.8 (M - H)\ 95% pure. Example 84. Preparation of Nα-isobutyI-Nα-(4-methylbenzenesulfonyl)-Ne-(N,α-t'ert- butoxycarbonyl-O-benzyl-L-threonyl)-L-lysine
The title compound was prepared from Nα-isobutyl-Nα-(4-methylbenze!nesulfonyl)-L-lysine (100 mg, 0.29 mmol, example 1, step E) as described in general procedure Be using commercially available Nα-tert-butoxycarbonyl-O-benzyl-L-threonine (93 mg, 0.3 mmol). The final product was triturated with ether to yield 195 mg (98%) of the desired material.
LC-MS : 646.9 (M - H)\ 95% pure.
Example 85. Preparation of Na-isoburyl-Na-(4-methylbenzenesulfonyl)-Ne-(N'a-tert- butoxycarbonyl-Nτ-benzyl-L-histidinyl)-L-lysine
The title compound was prepared from Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine (100 mg, 0.29 mmol, example 1, step E) as described in general procedure Be using commercially available Λ^α-tert-butoxycarbonyl-Nτ-benzyl-L-liistidine (100 mg, 0.3 mmol). The final product was triturated with ether to yield 133 mg (65%) of the desired material.
LC-MS: 682.2 (M - H)-, 90% pure.
Example 86. Preparation of Nα-isobutyl-Nα-(4-methyIbenzenesulfonyl)-Ne-(N,α-tert- butoxycarbonyI-L-tryptophanyl)-L-lysine
The title compound was prepared fromNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine (100 mg, 0.29 mmol, example 1, step E) as described in general procedure Be using commercially available Nα-tert-butoxycarbonyl-L-tryptophan (90 mg, 0.3 mmol). The final product was triturated with ether to yield 157 mg (81%) of the desired material.
LC-MS: 641.8 (M - H)-, 90% pure. • ' Example 87. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-(N'α-t:ert- butoxycarbonyl-O-benzyl-L-tyrosyl)-L-lysine
The title compound was prepared fromNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine (100 mg, 0.29 mmol, example 1, step E) as described in general procedure Be using commercially available Nα-tert-butoxycarbonyl-O-benzyl-L-tyrosine (111 mg, 0.3 mmol). The final product was triturated with ether to yield 145 mg (68%) of the desired material.
LC-MS: 708.2 (M - H)-, 95% pure.
Example 88. Preparation of Nα-isobutyl-Nα-(4-methyIbenzenesulfonyl)-Ne-(N'α-methyl-L- phenylalanyl)-L-lysine
This product 'was prepared from Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-(N'α-tert- butoxycaι-bonyl-NJά-methyl-L-phenylalanyl)-L-lysine (100 mg, 0.16 mmol, example 81) as described in general procedure C. The residue was triturated with ether and placed under high vacuum to yield a hard white foam (25 mg, .30%).
LC-MS: 516.3 (M -1H)\ 95% pure. '
Example 89. Preparation of Nα-isobutj'l-Nα-(4-methylbenzenesuifonyl)-Ne-(L-methionyl)- L-lysine
This product was prepared from Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-(N'α-tert-- butoxycarbonyl-L-methionyl)-L-lysine (95 rrig, 0.16 mmol, example 82) as described in general procedure C. The residue was triturated with ether and placed under high vacuum to- yield a hard white foam (40 mg, .51 %) .
LC-MS: 486.2 (M - H)-5 95% pure. , Example 90. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-[S-(4- methylbenzyl)-L-cysteinyl]-L-lysine
This product was prepared from Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-[N'α-tert- butoxycarbonyl-S-(4-methylbenzyl)-L-cysteinyl]-L-lysine (91 mg, 0.16 mmol, example 83) as described in general procedure C. The residue was triturated with ether and placed under high vacuum to yield a hard white foam (60 mg, 66%>).
LC-MS: 562.2 (M- H)\ 85% pure.
Example 91. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-(O-benzyl-L- threonyl)-L-lysine
This product was prepared from Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-(N'α-tert- butoxycarbonyl-O-benzyl-L-threonyl)-L-lysine (110 mg, 0.18 mmol, example 84) as described in general procedure C. The residue was triturated with ether and placed under high vacuum to yield a hard white foam (40 mg, 41%).
LC-MS: 549.3 (M - H)\ 95% pure.
Example 92. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-(Nτ-benzyl-L- histidinyl)-L-lysine
This product was prepared from Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-(N'α-tert- butoxycarbonyl-Nτ-benzyl-L-histidinyl)-L-lysine (110 mg, 0.16 mmol, example 85) as described in general procedure C. The residue was triturated with ether and placed under high vacuum to yield a hard white foam (51 mg, 54%).
LC-MS: 582.1 (M - H)-, 95% pure. , Example 93. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-(L- tryptophanyl)-L-lysine
This product was prepared from Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-(N'α-tert- butoxycarbonyl-L-tryptophanyl)-L-lysine (100 mg, 0.15 mmol, example 86) as described in i general procedure C. The residue was triturated with ether and placed under high vacuum to yield a hard white foam (27 mg, 33%):
LC-MS: 541.6 (M - H)\ 90% pure.
Example 94. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-(O-benzyl-L- tyrosyl)-L-lysine
This product was prepared from Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-(N'α-tert- butoxycarbonyl-O-benzyl-L-tyrosyl)-L-lysine (115 mg, 0.16 mmol, example 87) as described in general procedure C. The residue was triturated with ether and placed under high vacuum to yield a hard white foam '(19 mg, 19%).
LC-MS: 608.2 (M - H)\ 90% pure. . '
Example 95. Preparation of (2S,2'S) 2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N- (7Vα-benzoyl-S-pbenyrlalanyl)-2,6-diaminohexanol
The product of example 70, Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-(N'α-benzoyl-L- phenylalanyl)-L-lysine methyl ester, was reduced with LiAlH4 following the indications of genera] procedure G. The final product was obtained in 76% yield.
ΗΝMR (DMSO-d6): δ 0.77 (t,7= J0,6H), 0.90-1.30 (m.5H), 1.48 (m, IH), 1.85 (m, 1H), 2.38 (s, 3H), 2.55-2.70 (m, 3H), 2.75-2.85 (m, 2H), 2.93 (dd, 7= 13.5, 7.5, IH), 3.35 (m, IH), 3.50 (m, IH), 3.86 (m, IH), 4.65 (m, IH), 7.10 (d, 7= 7.8, 2H), 7.12-7.22 (m, 6H), 7.35 (d, 7= 8.0, 2H), 7.50 (d, 7= 7.8, 2H), 7.67 (d, 7= 8.2, 2H), 7.72 (t, 7- 5.0, IH), 7.70 (d, 7= 8.0, IH). Example 96. Preparation of (2S,2»S) 2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N- [7Vα-(4-methyIbenzenesuIfonyl)-S-phenylalanyl]-2,6-diaminohexanol
The product of example 68, Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-[N'α-(4- methylbenzenesulfonyl)-L-phenylalanyl]-L-lysine methyl ester, was reduced with LiAlH4 following the indications of general procedure G. The final product was obtained in 76% yield.
!HΝMR(DMSO-d6): δ 0.83 (t,7= 7.0, 6H), 0.90-1.30 (m, 5H), 1.52 (m, IH), 1.90 (m, IH), 2.36 (s, 3H), 2.80 (<id, 7= 1:2.0, 8.0,' IH), 2.85-3.10 (m, 5H), 3.52 (m, IH), 4.66 (m, 2H), 7.10-7.28 (m, 3 H), 7.30-J55 (m, 7H), 7.67 (d, 7= 8.0, 2H), 7.80 (d, 7= 8.0, 2H), 7.96 (m, IH), 8.51 (d, 7= 8.0, IH). '' ! • ; ■
Example 97. Preparation of (2S,2'S) 2-N-isobutyl-2-Λr-(4-methylbenzenesulfonyl)-6-N- [N'α-(4~methylbenzenesulfonyl)-S-tryptophanyl]-2,6-diaminohexanol
The product of ' example 69, Nα-is butyl-Nα-(4-methylbenzenesulfonyl)-Ne-[N'α-(4- methylbenzenesulfonyl)-L-tryptophanyI]-L-lysine methyl ester, was reduced with LiAlH4 following the indications of general procedure G. The final product was obtained in 65% yield.
'H ΝMR (DMSO-d6): δ θ.82 (d, 7= 7.0, 3H), 0.85 (d, 7= 6.8, 3H), 0.88-1.20 (m, 5H), 1.45 (m, IH), 2.30 (s, 3H), 2.36 (s, 3H), 2.62 (m, 2H), 2.76 (m, 2H), 2.90 (m, 2H), 2.90 (m, 2H), 3.20-3.40 (m, 2H), 3.50 (m, 1H)S 3.85 (m, IH), 4.67 (t, 7= 5.0, IH), 6.90 (t, 7= 7.4, IH), 7,03 (m, 2H), 7.13 (d, 7= 7.6, 2H), 7.27 (d, 7= 7.6, 2H), 7.35 (m, 3H), 7.46 (d, 7= 7.6, 2H), 7.68 (m, 3H), 7.82 (d, 7= 8.8, IH), 10.70 (s, IH).
Example 98. Preparation of Nα-isobutyl-Nα-(4-methyIbenze9esulfonyl)-Ne-[N'α-(4- methylbenzenesulfonyl)-L-phenylalanyl-N-cyanoamidine]-L-lysine
To a stirred solution of the thioamide, Nα7isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-[N,α-(4- methy]benzenesulfonyl -L-thiophenylalanyl]-L-lysine, (227 mg, 0.33 mmol, example 72) in MeOH (3 mL) was added cyanamide (28 mg, 0.66 mmol). The mixture was stirred for 5 min, then mercuric acetate (209 mg, 0.66 mmol) was added. The reaction was stirred for 3 h then diluted with saturated NH4C1 and extracted with EtOAc. The organic layer was washed with brine and concentrated then rediluted with THF/MeOH (2 nιL/1 mL) and treated with IN NaOH (0.8 mL). After stirring for 4 h, the reaction was acidified with IN HCl and extracted with EtOAc. The organic layer was dried over MgSO4, concentrated and purified by column chromatography to give 128 mg (57%) of the final product.
'HNMR (DMSO-d6): δ 0.80 (d, 7= 6.5, 3H), 0.83 (d, 7= 6.8, 3H), 1.00-1.12 (m, 4H), 1.35 (m, IH), 1.71 (m, IH), 1.88 (m, IH), 2.37 (s, 3H), 2.50 (s, 3H), 2.60-2.80 (m, 4H), 2.90 and 2.97. (ABX,7= 13.2, 7.0, 2H), 3.88 (m, IH), 4.15 (t,7= 7.0. IH), 7.10 (d, 7= 8.2, 2H), 7.13-7.23 (m, 5H), 7.38 (d, 7= 8.2, 2H), 7.49 (d, 7= 7.9, 2H), 7.68 (d, 7= 8.2, 2H), 7.78 (m, IH), 7.92 (d, 7= 8.3, IH), 12.70 (br s, IH). ■
Example 99. Preparation o Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-(N,α-acetyl-L- tryptophanyI)-L-lysine
Step A. Preparation of Nα-acetyl -L-tryptophan
L-tryptophan was reacted with acetyl chloride under the conditions used in general procedure A giving the title compound which was recrystallised from DCM (65%).
Η ΝMR (CDC13): δ 1.92 (s, 3H), 3.11-3.46 (m, 2H), 4.73 (t, 7= 4.5, IH), 6,92-7.10 (m, 3H), 7.28 (d, 7= 6.0, IH), 7.55 (d, 7= 6.0, IH). • '
Step B. Preparation , ■ of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-(N'α-acetyl-L- tryptophanyl)-L-lysine [ ■ ■ '
The title compound was prepared from Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine hydrochloride (400 mg, 1.0 mmol, example 1 , step E) suspended in THF (20 mL) and IN ΝaOH (0.5 mL, to reach pH 10) as described in general procedure Be using Nα-acetyl-L-tryptophan (300 mg, i .4 mmol) prepared in step A of this example, N-hydroxysuccinimide (115 mg, 1.0 mmol) and DCC (210 mg, 1.0 mmol). The crude material (600 mg) was purified by preparative HPLC to yield 411 mg (45%>) of the desired material.
Η NMR (CDC13): δ 0.82 (d57= 6.8, 3H), 0.84 (d, 7= 6.8, 3H), 1.08-1.11 (m, 2H), 1.18-1.22 (m,2H), 1.45-1.52 (m, lH), 1.94 (s, 3H),2.32 (s, 3H), 2.84-3.09 (m, 4H), 3.15-3.18 (m, lH), 4.19 (t, 7= 7.5, IH), 4.57 (t,7= 7.1, IH), 6.85 (t, 7= 7.2, IH), 7.04 (s, 2H), 7.32 (d, 7= 8.1, 2H), 7.56 (d, 7= 8.1, IH), 7.69 (d, 7= 7.2, 2H). LC-MS: 583.8 (M - H)-, 99% pure. .
Example 100. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-(N'α- pivaloyl-L-tryptophanyl)-L-lysine
Step A. Preparation of Nα-pivaloyl-L-tryptophan , '
L-tryptophan was reacted with pivaloyl chloride under the conditions used in general procedure A giving the title compound which was recrystallised from DCM (50%).
Η ΝMR (CDC13): δ 1.02' (s, 9H), 3.21-3.46 (m, 2H), 4.73 (t, 7= 4.5, IH), 6.92-7.10 (m, 3H), 7.28 (d, 7= 6.6, IH), 7.55 (d, 7= 6.0, IH). ;
Step B. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-(N'α-pivaloyl-L- tryptophanyl)-L-lysine •
The title' compound was prepared from Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine hydrochloride (400 mg, 1.0 mmol, example 1, step E) suspended in THF (20 mL) and IΝΝaOH (0.5 mL, to reach pH 10) as described in general procedure Be using Nα-pivaloyl-L-tryptophan (350 mg, 1.5 mmol) prepared in step A of this example, N-hydroxysuccinimide (115 mg, 1.0 mmol) and DCC (210 mg, 1.0 mmol). The crude material (488 mg) was purified by preparative HPLC to yield 244 mg (50%) of the desired material.
!HΝMR (CDCI3): δ 0.82(d,7= 6.8, 3H), 0.84.(d, 7= 6.8-, 3H), 1.06 (s, 9H), 1.08-1.11 (m, 2H)s 1.18-1.22 (m, 2H), 1.45-1.52 (m, IH), 2.38 (s, 3H), 2.84-3.09 (m, 4H), 3.15-3.18 (m, 1H), 4.24 (t, 7= 7.5, IH), 4.57 (t, 7= 7.1, IH), 6.85 (t, 7= 7.2, IH), 7.04 (s, 2H), 7.32 (d,7= 8.1, 2H), 7.56 (d, 7= 8.1 , IH), 7.69 (d, 7= 7.2, 2H). LC-MS: 625.8 (M - H)\ 99% pure.
Example 101: Preparation of Nα-isobutyl-Nα-(4-methyIbenzenesulfonyl)-N€-(N'α- trifluoroacetyl-L-phenylalanyl)-L-lysine
Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-(N'α-tert-butoxycarbonyl-L-phenylalanyl)-L- lysine (50 mg, 0.08 mmol, example 78) is added to a mixture of of trifluoroacetic anhydride and TFA and was stirred for a period of 30 min. Afterwards, the reactant were evaporated and the residue triturated with ether to give 40 mg (74%) of the title compound.
LC-MS: 598J (M - H)\ 99% pure.
Example 102. Preparation of (2S,2'S) 2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N- [7Vα-(4-methylbenzenesulfonyl)-S-thiotryptophanyl]-2,6-diaminohexanol
The product of example 75, Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-[N'α-(4- - methylbenzenesulfonyl)-L-thiotryptophanyl]-L-lysine methyl ester, was reduced with LiAlH4
•' i following the indications of general procedure G. The final product was obtained in 61% yield.
ΗΝMR (CDC13): δ.0.83 (d, 7= 7.0, 3H), 0.85 (d, 7= 7.0, 3H), 0.88-1.25 ( , 5H), 1.50 ( , IH), 1.88 (m, IH), 2.30 (s, 3H); 2.37 (s, 3H), 2.80 and 2.95 (ABX, 7= 14.5, 7.3, 2H), 2,90 and 3.10 (ABX, 7= 14.5, 7.8, 2H), 3.05 (m, 2H), 3.20-3.40 (m, 2H), 3.51 (m, IH), 4.30 (m, IH), 4.68 (m, lH), 6.90 (t,7= 7.5, lH), 7.02 (t,7= 7.5, lH), 7.06 (s, IH), 7.09 (d,7= 8.0, 2H), 7.28 (d, 7= 7.7,
IH), 7.32-7:42 (m' ; 5H),J.70 (d, 7= 8.10, 4H), 10.71 (s, IH).
Example 103. Preparation of Nα-isobutyl-Nα-(4-methyIbenzenesulfonyl)-Ne-[N,α-(4- methylbenzenesulfonyl)rL-tryptophanyl-N-cyanoamidine]-L-lysine
To a stirred solution of thioamide Nα-isobutyl-Nα-(4-metiιylbenzenesulfonyl)-Ne-[N'α-(4- methylbenzenesulfonyl)-L-thiotryptophanyl]-L-lysine (240 mg, 0.33 mmol, example 73) in MeOH (3 mL) was added cyanamide (28 mg, 0.66 mmol). The mixture was stirred for 5 min, then mercuric acetate (209 mg, 0.66 mmol) was added. The reaction was stirred for 3 h then diluted with saturated NH4C1 and extracted with EtOAc. The organic layer was washed with brine and concentrated then rediluted with THF/MeOH (2 mL/1 mL) and treated with IN NaOH (0.8 mL). After stirring for 4 h, the reaction was acidified with IN HCl and extracted with EtOAc. The organic layer was dried over MgSO4, concentrated and, purified by column chromatography to give 154 mg (65%) of the final product.
'HNMR (DMSO-d6): δ 0.80 (d,7= 6.5, 3H), 0.83 (d, 7= 6.8, 3H), 1.00-1.14 (m, 4H), 1.38 (m, IH), lJ2(m, lH),T.88 (m, IH), 2.30 (s, 3H), 2.37 (s, 3H), 2J0 (dd, 7= 13.5, 7.0, lH), 2.92 (m, 3H), 3.87 ( , IH), 4.12 (t, 7= 7.0, IH), 6.90 (t, 7= 7.5, IH), 7.00 (t,7= 7.5, IH), 7.04 (s, IH), 7.13 (d, 7= 8.3, 2H), 7.28 (d, 7= 8.3, IH), 7.38 (m, 3H), 7.47 (d, 7= 8.0, 2H), 7.65 (d, 7= 8.0, 2H), 7.72 (t, 7= 5.0, IH), 7.85 (d, 7= 8.1, IH), 10.73 (s, IH), 12.70 (br s, IH).
Example 104. Preparation of Nα-isobutyI-Nα-(4-methyIbenzenesuIfonyI)-Ne-(N'α- benzenesulfonyl-L-tryptophanyl)-L-lysine 2,3-dihydroxypropyl ester
Λfø-isobutyl-Λ^
(170 mg, 0.25 mmol) dissolved in a solution of DMF (1 L) was treated with glycerol (100 μL, 1.37 mmol) and EDC (1Q0 mg, 0.5 mmol) and stirred overnight. Then, the solution was poured in 5% citric acid and extracted with EtOAc. The solvent was evaporated and the residue purified by preparative HPLC to give 15 Omg (79%) of the final product.
Η NMR (CDC13): δ 0.81 (d, 7= 6.3, 3H), 0.83 (d, 7= 6.3, 3H), 0.94-1.03 (m, 4H), 1.32-1.35 (m, IH), 1.46-1.49 (m,TH), 1.81-1.93 (m, 2H), 2.35 (s, 3H), 2.75 (br s, IH), 2.82-2.99 (m, 4H), 3.45 (br s, 2H), 3.65 (br s,'lH), 3.82 (br s, 2H), 4.21 (t, 7= 7.2, IH), 6.85 (t, 7= 4.5, IH), 7.00 (t, 7= 4.5, IH), 7.23-7.31 (m, 6H), 7.42 (t, 7= 4.5, IH), 7.60 (d, 7= 6.8, 2H), 7.73 (d, 7= 6.8, 2H). Example 105. Preparation of Nα-isobutyl-Nα-(4-methyIbenzenesulfonyl)-Ne-(N'α- phenylglycyl)-L-lysine
Step A. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-chloroacetyl-L-lysine
The title compound was prepared from Nα-isobutyl-Nα-(4-methyIbenzenesulfonyl)-L-lysme potassium salt (2.0 g, 5.1 mmol) suspended in a mixture of THF (20 mL), DIEA (1.5 mL, 8.6 mmol) and chloroacetyl chloride (0.8 mL, 10.1. mmol). The reaction mixture was stirred for a period of 30 min. Then, a solution of 2Ν HCl (20 mL) was added and the solution extracted with EtOAc (25 mL, 3X). The organic phase was dried with MgSO4, filtered and evaporated to a brown oil. The crude material was purified by flash chromatography using a solvent gradient from 39: 1 to 19: 1 CH2Cl2/MeOH. The product was isolated as.a yellowish oil (1 J g, 78% yield).
Rf = 0.43 (EtOAc/hexane, 9 :1). LC-MS: 464 (M + H)+, 90% pure.
Step B . Preparation: of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-iodoacetyl-L-lysine
To a solution of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-chloroacety -L-lysine (1 J g, 3.9 mmol) in dry acetone (20 L) was added Νal. (1.5 g, 10.0 mmol). The'resulting mixture was stirred at room temperature for a period of 24 h. The precipitate ΝaCl was filtered off and the solvent evaporated to an oil. The crude material was dissolved in CH2C12 to precipitate the remaining Νal which was filtered off. The filtrate was washed with a 5%> aqueous solution of ΝaHSO3 (10 L) and with water (25 mL, 3X). The organic solvent was dried, filtered and evaporated to a brown oil (1.98 g, 96% yield) which was used without further purification in the next step.
Η MR (DMSO-d6): δ 0.81 (d,7= 6.5, 3H), 0.83 (d,7= 6.3, 3H), 1.20 (m, 2H), 1.33 (m, 2H),
1.45 (m, IH), 1.81 (m, IH), 1.89 (m, IH), 2.38 (s, 3H), 2.95 (m, 4H), 3.60 (s, 2H), 4.18 (t, 7=
7.2, IH), 7.38 (d, 7= 7.6, 2H), 7.67 (d, 7= 7.6, 2H), 8.17 (t, 7= 5.0, IH), 12.70 (br s, IH). LC-
MS: 556 (M + H)4, 99% pure. Step C. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-(N'α-phenylglycyl)-L- lysine
The title compound was prepared from Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne- iodoacetyl-L-lysine (200 mg, 0.38 mmol, product of step B above) by following the indications, of general procedure H using DIEA (0.19 mL, 1.09 mmol) and aniline (0.16 mL, 1.76 mmol). The crude material was purified by preparative HPLC. The product Was isolated as a solid (90 mg, 48% yield). . ; ■
!HΝMR (DMSO-d6): δ 0.79 (d, 7= 6.7, 3H), 0.81 (ds7= 6.9, 3H), 1.15 (m, 2H), 1.32 (m, 2H), 1.41 ( , IH), 1.77 (m, IH), 1.88 (m, IH), 2.37 (s, 3H), 2.90-3.01 (m, 4H), 3.58 (d, 7= 3.1, 2H), 4.14 (1,7= 7.1, IH), 5.86 (s, lH), 6.52 (d,7= 7J, 2H), 6.57 (t,7= 7.1, IH), 7.08 (t, 7= 7.6, 2H), 7.36 (d, 7= 7.6, 2H), 7.66 (d, 7= 8.2, 2H),J.80 (t, 7= 5.0, IH), 12.5 (s, IH). LC-MS: 490 (M + H)+, 99% pure. . i
Example 106. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-[N'α-(3- : pyridyI)glycyl]-L-lysine
The title ' compound was prepared from Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne- iodoacetyl-L-lysine: (200 mg, 0.38 mmol, example 105, step B) by following the indications of general procedure H using DIEA (0.19 mL, 1.09 mmol) and 3-aminopyridine (175 mg, 1.36 mmol). The crude material was purified by preparative HPLC. The product was isolated as a
• ! ' ' ' i ' ■' , ' solid (42 mg, 22% yield). -
'H ΝMR DMSO-d : δ 0.75 (d, 7= 6.7, 3H), 0.81 (d, 7= 6.7, 3H), 1.23-1.37 (m, 4H), 1.4.6 (m, IH), 1.84 (m, IH), 1.99 (m, IH), 2.31 (s, 3H), 2.85 (m, 1H),.3.07 (m, 3H), 3.95 (t, 7= 6.6, IH), 5.21 (d, 7= 9.3, 2H), 7.01 (s, 2H), 7.26 (d, 7= 8.0, 2H), 7.55 (d, 7= 8.6, 2H), 7.62 (t, 7= 7.0, 1H), 7.75 (d, 7= 7.8, 2H), 8.03 (d, 7= 5.4, IH), 8.21 (s, 1H), 9.28 (t, 7= 4.8, IH). LC-MS:
I -
491 (M + H)+, 95% pure.
Ill Example 107. Preparation of Nα-isobutyl-Nα-(4-methyIbenzenesulfonyl)-Ne-[N'α-(2,3- dimethoxybenzyl)glycyl]-L-lysine
The title compound was prepared from Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne- iodoacetyl-L-l sine (150 mg, 0.29 mmol, example 105, step B) by following the indications of general procedure H using DIEA (0.14 mL, 0.81 mmol) and 2,3-dimethoxybenzylamine (0.12 mL, 0.81 mmol). The crude material was purified by preparative HPLC. The product was isolated as a solid (25 mg, 16% yield). • '
ΗΝMR(DMSO-d6): δ 0.79 (d,7= 6.9, 3H), 0.81 (d, 7= 6.9, 3H), 1.20 (m, 2H), .1.30-1.50 (m, 3H), 1.75-1.95 (m, 2H), 2.36 (s, 3H), 2.90-3.05 (m, 4H), 3.09 (s, 2H), 3.68 (s, 2H), 3.72 (s, 3H), 3.79 (s, 3H), 4.16 (t, 7= 6.6, IH), 6.95 (m, 2H), 7.01 (d, 7= 7.6, IH), 7.35 (d, 7= 7.9, 2H), 7.67 (d, 7= 8.0, 2H), 7.80 (t, 7= 5.4, IH). LC-MS: 564 (M + H)+, 98% pure.
Example 108. Preparation of Nα-isobutyl-Nα-(4-methyIbenzenesulfonyl)-Ne-[N'α-(2- pyridyl)glycyl] -L-lysine
The title compound was prepared from Nα-isobutyl-Nα-(4-methyIbenzenesulfonyl)-Ne- iodoacetyl-L-lysine (200 mg, 038 mmol, example 105, step B) by following the indications of general procedure H using DIEA (0.19 mL, 1.09 mmol) and 2-aminopyridine (170 mg, 1.31 mmol). The crude material was purified by preparative HPLC. The product was isolated as a solid (88 mg, 47% yield).
ΗΝMR (DMSO-d6): δ 0.79 (d, 7= 6.1, 3H), 0.82 (d,7= 6.4, 3H), 1.2 (m, 2H), 1.39-1.50 (m, 3H), 1.75 - 1.90 (m, 2H), 2.37 (s, 3H), 2.87-3.04 (m, 4H), 4.21 (t,7= 6.6, IH), 4.90 (s, 2H), 6.88 (t, 7= 6.5, IH), J05 (d, 7= 8.9, IH), 7.37 (d,7= 7.9, 2H), 7.6J(d, 7= 8.1, 2H), 7.88 (t, 7= 8.1, IH), 7.95 (d,7= 6.3, IH,), 8.33 (t,7~ 5.6 IH), 8.41 (s, 2H), 12.7 (s, IH). LC-MS: 491 (M + H)+, 99% pure. ■ Example 109. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-(N,α- benzylglycyl)-L-lysine
The title compound was prepared from Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne- iodoacetyi-L-lysine (200 mg, 0.38 mmol, example 105, step B) by following the indications of general procedure H using DIEA (0.20 mL, 1.10 mmol) and benzylamine (320 mg, 2.99 mmol). The crude material was purified by preparative HPLC. The. product was isolated as a solid (137 mg, 71% yield). ;
'H ΝMR'(CDC13): δ 0.81 (s, 6H), 1.26-1.60 (m, 5H), 1.80-2.05 (m, 2H), 2.40 (s, 3H), 2.90- 3.00 (m, 3H), 3.04 (s, IH), 3.19 (m, 2H), 3.8.7 (s, IH), 4.26 (s, 2H), 6.16 (s, 4H), 7.28 (d,7= 5.3, 2H), 7.38 (m, 3H), 7.48 '(d, 7= 4.5, 2H), 7.70 (d, 7= 7.5, 2H ), 8.90 (s, IH). LC-MS: 502 (M - H)\ 99% pure; ! ■ ' ' '
Example 110. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-[N'α- (piperidinyl-N-ethyl)glycyl]-L-lysine trifluoroacetic acid salt
The title compound was prepared from Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne- iodoacetyl-L-lysine (150 mg, 0.29 mmol, example 105, step B) by following the indications of general procedure H using DIEA (0.12 mL, 0.69 mmol) and l-(2-aminoethyl)piperidine (0.12 mL, 0.84 mmol). The crude material was purified by preparative HPLC. The product was isolated as a solid (65 mg, 33% yield). I
'H ΝMR (DMSO-d6): δ 0.80 (d, 7= 6.3, 3H), 0.82 (d, 7= 6.7, 3H), 1.24-2.00 ( , 13H), 2.38 (s, 3H), 2.85-3.40 (m, 12H), 3.76 (s, 2H), 4.21 (t, 7= 6.7, IH), 7.37 (d, 7= 7.9, 2H), 7.67 (d, 7 = 8.0, 2H), 8.49 (t, 7 « 5.0, IH), 9.25 (s, 2H). LC-MS: 525 (M + H)+, 99% pure. Example 111. Preparation of Nα-isobutyl-Nα-(4-methyIbenzenesuIfonyl)-Ne-[N'α-(4- morpholinyl-N-ethyl)glycyl]-L-lysine
i
The title compound was prepared from Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne- iodoacetyl-L-lysine (150 mg, 0.29 mmol, example 105, step B) by following the indications of general procedure H using DIEA (0.12 mL, 0.69 mmol) and 4-(2-aminoethyl)morpholine (0.11 mL, 0.84' mmol). The crude material was purified by preparative HPLC. The product was isolated as a solid (51 mg, 34% yield). ;
■ : ' 'i
]H ΝMR (DMSO-d6): δ 0.80 (d, 7= 6.1, 3H), 0.82 (d, 7= 7.0, 3H)J 1.24-1.30 (m, 2H), 1.35- 1.55 (m, 3H), 1.75-1.95 <m, 2H), 2.38 (s, 3H), 2.80-3.25 (m, 10H), 3.30 (s, 2H), 3.75 (s, 6H), 4.21 (t,7= 7.1, lH), 7.37 (d, 7= 8.3, 2H), 7.68 (d, 7= 8.4, 2H), 8.47 (m, IH). LC-MS: 525 (M - H)\ 99% pure.
Example 112. Preparation of Nα-isobutyl-Nα-(4-methyIbenzenesu.fonyI)-Ne-[N,α-(4- pyridyl)g!ycyl]-L-Iysine
The title compound was prepared from Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne- iodoacetyl-L-lysine (150 mg, 0.29 mmol, example 105, step B) by following the indications of general procedure H using DIEA (0.10 mL, 0.57 mmol) and 4-aminopyridine (76 mg, 0.59 mmol). The crude material was purified by preparative HPLC. The product was isolated as a solid (39 mg, 28% yield).
Η ΝMR (DMSO-d6): δ 0.79 (d, .7= 6.8, 3H), 0.81 (d, 7= 6.5, 3H), 1.25 (m, 2H), 1.35-1.51 (m, 3H), 1.78- 1.91 ( , 2H), 2.37 (s, 3H), 2.90 (m, IH), 3.00 (m, 3H), 4.20 (t, 7= 7.1, IH), 4.85 (s, 2H); 6.80 (d, 7= 6.7, 2H), 7.37 (d, 7= 8.2, 2H), 7.67 (d, 7= 7.6, 2H), 8.02 (d, 7= 7.0,
! i ' i
2H), 8.15 (s, 2H), 8.37 (t, 7= 4.9, IH), 12.74 (br s, IH). LC-MS: 489 (M - Hr, 99% pure. Example 113. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-[N,α-(3- quinolyl)glycyl]-L-lysine
The title compound was prepared from Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne- iodoacetyl-L-Iysine (200 mg, 0.38 mmol, example 105, step B) by following the indications of general procedure H using DIEA (0.19 mL, 1.09 mmol) and 3-aminoquinoline (260 mg, 1.80 mmoi). The crude material was purified by preparative HPLC. The product was isolated as a solid (24 mg, 12% yield).
]H ΝMR (DMSO-d6): δ 0.76 (d, 7= 5.9, 3H)„ 0.82 (d, 7= 6.5, 3H), 1.26-1.50 (m,.5H), 1.80- 2.10 (m, 2H), 2.34 (s, 3H), 2.86 (m, IH), 3;04-3.20 (m, 2H), 4.00 (m, IH), 5.67 (d, 7=4.8, 2H), 6.85 (s, 2H), 7.29 (d, 7= 7.8, 2H), 7.75 (m, 4H), 8.02 (s, IH), 8.07 (d, 7= 8.0, IH), 8.12 (d, 7 = 8.5, IH), 8.97 (m, IH), 9.05 (s, IH). LC-MS: 539 (M - H)", 95% pure.
Example 114. Preparation of Nα-isobutyI-Nα-(4-methylbenzenesulfonyl)-Ne-(N,α- benzyl-N'α-phenylglycy -L-lysine
Step A. Preparation of N-benzyl-N-phenyl gly cine
: Commercially available N-phenylglycine (1.51 g, 10.0 mmol) dissolved. in THF (20 mL) was treated with butyllithium 2.5M in hexane (8.8 mL, 22.0 mmol) under an inert atmophere of argon at' - 78 °C. The mixture was stirred for 15 min before the addition of benzylbromide (1.3 mL, 1 1 mmol). The reaction mixture was stirred for 2 h while warming up to room temperature.. Then, 2Ν HCl '(25 mL) was added and the mixture extracted with EtOAc (30 mL, 3X). The organic phase was dried over MgSO4, filtered and concentrated to an oil. The crude material was purified by flash chromatography using a solvent gradient from 19 : 1 to 9 : 1 CH2Cl2/MeOH. The product was isolated as a brown and hardy oil (1.98 g, 82%).
'H NMR (CDC13): δ 4.16 (s, 2H), 4.68 (s, 2H), 6.76 (d, 7= 8.5, 2H), 6.83 (t, 7= 7.2, IH), 7.27 (t, 7= 6 , 2H), 7.33 (m, 3H), 7.38 (t,-7= 7.6, 2H), 10.70 (s, IH). Step B. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-(N,α-benzyl-N'α- phenylgIycyl)-L-lysine
The title compound , was prepared from solid phase bound Nα-isobutyl-Nα-(4- methylbenzenesulfonyl)-Ne-(9-fluorenylmethoxycarbonyl)-L-lysine (200 mg) in asimilar fashion to general procedure Bb using N-benzyl-N-phenylglycine (100 mg, 0.45 mmol) prepared in step A of this example with DCC (200 mg, 0.97 mmol) and HOBt (100 mg, 0.74 mmol) as the activating reagents.' The final product was purified by preparative HPLC to yield 21 mg (75%>) of the desired material. ;
,HΝMR (DMSO-d6): δ 0J9 (d, 7= 6.9, 3H), 0.82 (d, 7= 7.2, 3H), 1.17 (m, 2H), 1.33 (m, 2H), 1.42 (m, IH), 1.77 (m, IH), 1.88 (m, IH), 2.37 (s, 3H), 2.90-3.01 (m, 4H), 3.94 (s, 2H), 4.16 (t, 7= 7.2, IH), 4.62 (s, 2H), 6.59 (m, 3H), 7.11 (t, 7= 7.5, 2H), 7.24 (m, 3H), 7.31 (t, 7= 7.3, 2H), 7.36 (d, 7= 7.6, 2H), 7.66 (d, 7= 8.5, 2H), 7.87 (t, 7= 5.3, IH), 12.6 (s, IH). LC-MS: 578 (M + H)+, 85% pure.
Example 115. Preparation of Nα-isobutj -Nα-(4-methylbenzenesulfonyl)-Ne-(N,α- methyl-N'α-phenylglycyl)-L-lysine
Step A. Preparation of N-methyl-N-phenylglycine
The title compound, was prepared from N-phenylglycine (200 mg, 1.3 mmol) as described for the preparation of N-benzyl-N-phenylglycine (example 114, step A) using methyl iodide (1 mL, 6.6 mmol) instead of benzylbromide. The crude material was purified by preparative HPLC. The product was isolated as a solid (90 mg, 41% yield).
]H ΝMR (CDC13): δ 3.07 (s, 3H), 4.09 (s, 2H), 6.73 (d, 7= 7.7, 2H), 6.80 (t, 7= 7.2, IH), 7.27
(t, 7= 7.3, 2H), 8.45 (s, IH).
)
Step B. • Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-(N'α-methyl-Λ α- phenylglycyl)-L-lysine ' . The title compound was prepared from solid phase bound Nα-isobutyl-Nα-(4- methylbenzenesulfonyl)-Ne-(9-fluorenylmethoxycarbonyl)-L-lysine (200 mg) ' in a similar fashion to general procedure Bb using N-methyl-N-phenylglycine (75 mg, 0.45.mmol) prepared in step A of this example with DCC (200 mg, 0.97 mmol) and HOBt (100 mg, 0.74 mmol) as the activating reagents. The final product was purified by preparative HPLC to yield 7 mg (25%o) of the desired material.
ΗΝMR(DMSO-d6): δ 0J9 (d, 7= 6.5, 3H), 0.82 (d, 7= 7.0, 3H), 1.16 (m, 2H), 1.32 (m, 2H), 1.41 (m, IH), 1.77 (m, IH), 1.88 (m, IH), 2.37 (s, 3H), 2.90-3.01 (m, .4H), 2.96 (s, 3H), 3.82 (s, 2H), 4.16 (t, 7= 7.0, IH), 6.62 (m, 3H), 7.16 (t, 7= 7.4, 2H), 7.36 (d, 7= 8.1, 2H), 7.66 (d, 7= 8.0, 2H), 7.80 (t, 7= 43, IH), 12.7 (s, IH). LC-MS: 502 (M + H)+, 98% pure.
Example 116. Preparation of (2S,2'S) 2-N-isobutyI-2-N-(4-methyIbenzenesulfonyl)-6-N- [7Vα-(4-methylbenzenesulfonyl)-S-tryptophanyl]-2,6-diaminohexanal
Step A. Preparation of (2S) 2-N-isobutyl-2-N-(4-methylbenz'enesulfonyl)-2,6-diaminohexanol
A solution ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine (12.0 g, 30.5 mmol, example 1 , step E) dissolved in MeOH (100 mL) was treated with of trimethylsilyl chloride (20 mL). The mixture was refluxed 5 h before stirring at room temperature for 5 h. Afterwards, the solution was evaporated and placed under high vacuum until a hard foam was obtained (13.6 g). This was dissolved in dry THF (100 mL) and added dropwise to a solution of LiAlH4 (5.0 g, 131.6 mmol) in THF (300 L). The solution was stirred for 2 h, then heated to reflux for 20 min. After cooling in an ice bath the solution was quenched by addition of MeOH (5 mL), water (5 mL), then 10%) ΝaOH (5 mL). The solvent was evaporated and the precipitate dissolved in MeOH (200 mL) was stirred for 2 h at 60 °C. The granular precipitate formed was filtered off through celite, and the liquor was concentrated to form a clear oil which solidified on standing (8.48 g, 81 %). 'H NMR (CDCI3): δ 0.85 (d, 7= 63, 3H), 0.90 (d, 7= 6.3, 3H), 1.00-1.08 (m, IH), 1.29-1.34 (m, 3H), 1.92-1.97 (m, lH), 2.35 (s, 3H), 2.50 (t, 7= 6.7, 2H), 2.80-3.09 (m, 4H), 3.52 (d, 7= 5.1, 2H), 3.60-3.62 (t, 7= 6.8, IH), 7.22 (d, 7= 7.8, 2H), 7.63 (d, 7= .7.8, 2H).
Step B. Preparation of (2S,2'S) 2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N-[N'α-(4- methylbenzenesulfonyl)-S-tryptophanyl]-2,6-diaminohexanol
The title product was prepared from (2S) .2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6- diaminohexanol (400 mg, 1.2 mmol, step A) following the indications of general procedure Bd using Nα-(4-methylbenzenesulfonyl)-L-tryptophan (450 mg, 1.3 mmol, example 2, step A), EDC (290 mg, 1.5 mmol) and HOBt (100 mg, 1.5 mmol) . The crude material was purified by flash chromatography using a solvent gradient from 39:1 to 19:1 CH2Cl2/MeOH. The product was isolated as a yellowish oil (420 mg, 48% yield).
Η ΝMR (CDC13): δ 0.93 (t, 7= 6.8, 6H), 0.98 ( , IH), 1.26 (m, 3H), 1.42 (m, IH), 1.93 ( , 4H), 2.38 (s , 3H), 2.42! (s, 3H), 2.91 (m, IH), 3.05 (m , 3H), 3.13 (m, 2H), 3.56 (m, 2H), 3.66 (m, IH), 3.91 (d, 7= 5.6, IH), 5.08 (d, 7= 5.9,1H), 6.32 (t, 7= 4.1, 1H),<6.95 (s, IH), 7.02 (t, 7 = 7.5, IH), 7.12 (d,7= 7.4, 2H), 7.19 (1, 7= 7.5, IH), 7.29 (d,7= 8.6, 2H), 7.35 (d,7= 7.8, IH), 7.51 (d, 7= 7.3, 2H), 7.72 (d, 7= 7.9, 2H), 8:54 (s, IH). LC-MS: 681 (M - H)\ 98% pure.
Step C. Preparation of (2S,2'S) 2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N-[Nα-(4- methylbenzenesulfonyl)-S-tryptophanyl]-2,6-diaminohexanal
The title compound was prepared from the previous derivative (400 mg, 0.59 mmol) by oxidation under Swern's reaction conditions using oxalyl chloride (110 μL, 1.2 mmol), DMSO (166 μL, 2.4 mmol), and triethylamine (2 mL) (Organic Syntheses, Collective Volume VII, p. 258-263). The crude material was purified by flash chromatography using a solyent gradient from 39.T to 19:1 CH2Cl2/MeOH. The product was isolated as a yellowish solid (330 mg, 83% yield).
]H ΝMR (DMSO-d6): δ 0.82 (t, 7= 7.6, 6H), 0.92 (m, IH), 1.09 (m, IH), 1.31.(m, 3H), 1.72 (m, IH), 1.83 (m, IH), 2.33 (s , 3H), 2.37 (s, 3H), 2.73 (d, 7= 4.8, IH), 2.96 (m, 5H), 4.08 (t, 7 = 6.1, IH), 4.72 (m, IH), 6.93 (t, 7= 7.4, IH), 6.99 (t, 7= 7.3, IH), 7.15 (d, 7= 7.6, IH), 7.37 (m, 6H), 7.61 (d, 7= 7.5, IH), 7.72 (d, 7= 7.6, 2H), 8.02 (t, 7= 5.7, IH), 9.51 (s,TH), 11.35 (s, IH).
Example 117. Preparation of (2S,TS) 2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N- (7Vα-pivaloyl-S-tryptophanyl)-2,6-diaminohexanol
The title product was prepared from (2S) 2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6- diaminohexanpl (example 116, step A) following the indications of general procedure Bd using Nα-pivaloyl-L-tryptophan (example 100, step A). Purification by HPLC gave 85 mg (48%) of the desired material.
'H ΝMR (CDCI3): δ 0.90-0.98 (m, 6H), 1.09-1.16 (m, IH), 1.23 (s, 9H), 1.25-1.29 (m, IH), 1.45-1.52 ( ,"lH), 1.92-1.97 (m, IH), 2.38 (s, 3H), 2.82-3.10 (m, 4H), 3.11-3.26 (m, 3H), 3.51- . 3.55 (m, IH), 3.51 (d, 7= 7.0, 2H), 4.64 (br s, IH), 6.97-7.21 (m, 6H), 7.35 (q, 7= 6.7, IH), 7.61 (d, 7= 7.1, IH), 7.69 (d, 7= 7.6, IH). LC-MS: 611.2 (M - H)\ 95% pure.
Example 118. Preparation of (2S,2'S) 2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N- (7Vα-acetyl-S-phenylalanyl)-2,6-diaminohexanol
The title product was prepared from (2S) 2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6- diaminohexanol (example 116, step A) following the indications of general procedure Bd using Nα-acetyl-L-phenylalanine (example 30, step A). Purification by HPLC gave 82 mg (51%o) of the desired material. ' '■ '•
'H ΝMR (CDCI3): δ 0.79 (d, 7= 6.3, 3H), 0.82 (d, 7= 6.3, 3H), 1.08-1.11 (m, 2H), 1.23-131 (m, 2H), 1.45-1.52 (m, IH), 1.92-1.97 (m, IH), 1.98 (d, 7= 9.0, 3H), 2.38 (s, 3H), 2.84-3.09 (m, 6H), 3.51 (d, 7= 7.1, 2H), 3.61-3.64 (m, 1H), 4.51 (q, 7= 6.9, IH), 7.09-7.26 (m, 7H), 7.73 (d, 7= 8.1, 2H). LC-MS: 530.4 (M - H)\ 99% pure. Example 119. Preparation of (2S,2'S) 2-N-isoburyl-2-N-(4-methylbenzenesulfonyl)-6-N- (7Vα-pivaloyl-S-phenylalahyl)-2,6-diaminohexanol
The title product was prepared from (2S) 2-N-isobutyl-2-N-(4-methylbenzenesul'fonyl)-2,6- dia inohexanol (example 116, step A) following the indications of general procedure Bd using Nα-pivaloyl -L-phenylalanine (example 52, step A). Purification by HPLC gave 65 mg (37%) of the desired material.
'HΝMR DC J: δ 0J9 (d,7= 63, 3H), 0.82 (d,7= 63, 3H), 1.08-1.11 (m, 2H), 1.13 (s, 9H), 1.23-1.31 (m,' 2H), 1.45-1.52 (m, IH), 1.92-1.97 (m, IH), 2.38 (s, 3H), 2.84-3.09 (m, 6H), 3.51 (d, 7= 7.1, 2H), 3.2 (t, 7= 6.1, IH), 4.51 (q, 7= 6.9, IH), 7.09-7.26 (m, 7H), 7.73 (d, 7= 7.9, 2H). LC-MS: 572.2 (M - H)\ 99% pure.
Example 120. Preparation of (2S,2'S) 2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N- [7V1α-(4-morpholinecarbonyl)-S-phenylaIanyl]-2,6-diaminohexanol
The title product was prepared from (2S) 2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6- diaminohexanol (example 116, step A) following the indications of general procedure Bd using Λrα-(4-morpholinecarbonyl)-L-phenylalanine (example 51, step A). Purification by HPLC gave 69 mg (38%>) of the desired material.
Η ΝMR (CDC13).: δ 0.79 (d, 7= 6.3, 3H), 0.82 (d, 7= 6.3, 3H), 1.08-1.11 (m, 2H), 1.23-131 (m, 2H), 1.45-1.52 (m,' IH), 1.92-1.97 (m,. lH), 2.39 (s, 3H), 2.84-3.09 (m, 6H), 3.21-331 (m, 4H), 3.45-3.64 (m, 7H), 4.48 (br s, IH), 7.09-7.26 (m, 7H), 7.73 (d, 7= 8.1, 2H). LC-MS: 601.1 (M - H)-, 99% pure.
Example 121. Preparation of (2S,2'S) 2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N- [7Vα-(4-acetylaminobenzenesulfonyl)-iS'-phenylalanyl]-2,6-ό')iaminohexanol
The title product was prepared from (2S) 2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6- diaminohexanol (example 116, step A) following the indications of general procedure Bd using Nα-(4-acetamidobenzenesulfonyl)-L-phenylalanine (example 3, step A). Purification by HPLC gave 103 mg (49%) of the desired material. .
ΗΝMR (CDC13): δ 0J9 (d, 7= 63, 3H), 0.82 (d, 7= 63, 3H), 1.08-1.11 (m, 2H), 1.23-131 (m, 2H), 1.45-1.52 (m, IH), 1.92-1.97 (m,. lH), 2.20 (s, 3H), 2.38 (s, 3H), 2.85-3.02 (m, 6H), 3.51 (^ 7= 7.1, 2^, 3.64 (^ 7= 6.4, IH), 3.88 (q, 7= 6.9, IH), 6.89'(br s, 2H), 7.19 (br s, 3H), 7.24 (d, 7= δ'.O, 2H), 7:54-7.61 (m, 4H), 7,73 (d, 7= 8.1, 2H). LC-MS: 685.2 (M - H)\ 99% pure. ■, : ' ' ■ ' '
Example 122. Preparation of (2S,2'S) 2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N- [7Vα-(2-thiophenesulfonyl)-S-phenylalanyl]-2,6-diaminohexanol
The title product was prepared from (2S) 2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6- diaminohexanol (example 116, step A) following the indications of general procedure Bd using Nα-(2-thiophenesulfonyl)-L-phenylalamne (example 24, step A). Purification by HPLC gave 69 mg (36%) of the desired material.
'H MR (CDC13): δ 0J9 (d, 7= 6.3, 3H), 0.82 (d, 7= 6.3, 3H), 1.08-1.11 (m, 2H), 1.28-135 (m, 2H), 1.45-1-52 (m, IH), 1.92-1.97 (m, IH), 2.39 (s, 3H), 2.85-3.18 (m, 6H), 3.51 (d, 7= 7.0,' 2H), 3.64 (q, 7= 6.4, IH), 3.94 (q, 7= 6.9, IH), 6.91-6.98 (m, 3H), 7.19 (br s, 3H), 7.24 (d, 7= 8.0, 2H), 7.35 (s, IH), 7.52 (d, 7= 4.1, IH), 7.73 (d, 7= 8.1, 2H). LC-MS: 634.2 (M - H)", 99% pure. • '
Example 123. Preparation of (2S,2'S) 2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N- (7Vα-benzenesulfonyl-S-phenylalanyl)-2,6-diaminohexanol
The title product was prepared from (2S) 2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6- diaminohexanol (example 116, step A) following the indications of general procedure Bd using •
Nα-benzenesulfonyl-L-phenylalanine (example 8, step A). Purification by HPLC gave .68 mg (36%>) of the desired material. ΗNMR(CDC13): δ 0.79 (d, 7=6.3, 3H), 0.82 (d, 7=6.3, 3H), 1.08-1.11 (m,2H), 1.28-135 (m,2H), 1.45-1.52 (m, IH), 1.92-1.97 (m, 1H),2.41 (s,3H), 2.85-3.18 (m,6H), 3.51 (d,7=7.0, 2H), 3.64 (q, 7= 6.4, IH), 3.84 (q, 7= 6.9, IH), 6.88 (d, 7= 6.6, 2H), 7.14-7.19 (m, 3H), 7.24 (d, 7= 8.0, 2H), 7.35 (1,7= 6.8, 2H), 7.52 (t,7= 7.1, IH), 7.60 (d,7= 6.8, 2H), 7J2 (d,7= 8.0, 2H). LC-MS: 628.2 (M-H)\ 99% pure. , :
Example 124. Preparation of (2S, S) 2-N-isobutyl-2-N-(4-methyIbenzenesulfonyI)-6-N- [N'α-(4-methylbenzenesulfonyl)-S-4-nitrophenylalanyl] -2,6- diaminohexanol
The title product was prepared from (2S) 2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6- diaminohexanol (example 116, step A) following the indications of general procedure Bd using Λrα-(4-methylbenzenesulfonyl)-L-4-nitrophenylalanine (example 26, step A). Purification by HPLC gave 101 mg (48%>) of the desired material.
'HΝMR (CDC13): δ 0.89-0.97 ( , 6H), 1.18.1.21 (m, 2H), 135-1.45 (m, 3H), 1.92-1.97 (m, ■lH),2.33,(s,3H), 239 (s,3H), 2.85-3.01 (m,3H), 3.15-3.24 (m,3H), 3.51 (d,7=7.0,2H),3Jl (q,7=6.4,lH),3.98(brs,lH),7.01(d,7=7.6,2H),7.19(d,7=8.0,2H),7.24(d,7= 8.0, 2H), 7.39 (d, 7= 7.7, 2H), 7.73 (d, 7= 8.1, 2H), 7.88 (d, 7= 7.9, 2H). LC-MS: 687.8 (M - H)", 99% pure. ;
Example 125. Preparation of (2S,2'S)2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N- [7Vα-(4-fluorobenzenesulfonyl)-S-phenylalanyl]-2,6-diaminohexanol
The title product was prepared from (2S) 2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6- diaminohexanol (example 116, step A) following the indications of general procedure Bd using Nα-(4-fluorobenzenesulfonyl)-L-phenylalanine (example 18, step A). Purification by HPLC gave 66 mg (34%) of the desired material.
]HΝMR(CDC13): δ 0.79 (d, 7=6.3, 3H), 0.82 (d, 7=6.3, 3H), 1.09-1.16 (m,2H), 135-139 (m, 2H), 1.45-1.52 (m, IH), 1.92-1.97 (m, IH), 2.42 (s, 3H), 2.82-3.18 (m, 6H), 3.51 (d,7= 7.0, 2H), 3.64 (q, 7= 6.3, IH), 3.79 (q, 7= 6.9, IH), 6.91 (d, 7= 7.0, 2H), 7.00 (t, 7= 6.8, 2H), 7.15- 7.21 (m, 3H), 735 (s, lH), 7.52 (d,7=7.1, lH), 7.54 (q, 7= 6.6, 2H), 7.73 (d, 7= 8.1, 2H). LC-, MS: 647.6 (M - H)-, 99% pure. ' ' '
Example 126. Preparation of (2S,2'S) 2-N-isoburyl-2-N-(4-methylbenzenesulfonyl)-6-N- [7Vα-(4-methoxybenzenesulfonyl)-S-phenylalanyl]-2,6-diaminohexanol
Step A. Preparation of Nα-(4-methoxybenzenesulfonyl)-L-phenylalanine
L-phenylalanine was reacted with 4-methoxybenzenesulfonyl chloride under the conditions used in general procedure A giving the title compound which was used without purification in the next step.
Step B. Preparation of (2S,2'S) 2-N-isobutyl-2-N-(4-methylberrzenesulfonyl)-6-N-[Nα-(4- methoxybenzenesulfonyl)-S-phenylalanyl]-2,6-diaminohexanol
The title product was prepared from (2S) 2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6- diaminohexanol (example 116, step A) following the indications of general procedure Bd using Nα-(4-methoxyberrzenesulfonyl)-L-ρhenylalanine (this example, step A). Purification by HPLC gave 107 mg (53%) of the desired material.
]H ΝMR (CDC13): δ 0J9 (d, 7= 6.3, 3H), 0.82 (d, 7= 6.3, 3H), L09-1.16 (m, 2H), 135-139 (m, 2H), 1.45-1.52 (m,lH), 1.94-1.97 (m, IH), 2.40 (s, 3H), 2.82-3.18 (m, 6H), 3.51 (d, 7= 7.0, 2H), 3.64 (q, 7= 6.3, IH), 3.79 (q, 7= 6.9, IH), 3.92 (s, 3H), 6.87 (d, 7= 6.9, 2H), 6.89 (d, 7 = 7.0, 2H), 7.15-721 (m, 3H), 7.35 (d, 7= 7.1, 2H), 7.50 (d, 7= 7.1, IH), 7.73 (d, 7= 8.1, 2H). LC-MS: 658.3 (M - H)", 99% pure. Example 127. Preparation of (2S,TS) 2-N-isobutyl-2-N-(4-methyIbenzenesuIfonyI)-6-N- [N'α-(4-trifluoromethylbenzenesulfonyl)-S-phenylalanyl] -2,6- diaminohexanol
The title product was prepared from (2S) 2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6- diaminohexanol (example 116, step A) following the indications of general procedure Bd using Nα-(4-trifluoromethylbenzenesulfonyl)-L-phenylalanine (example 23, step A). Purification by HPLC gave 49 mg (23%) of the desired material.
!H ΝMR (CDCI3): δ 0J9 (d, 7= 6.3, 3H), 0.82 (d, 7= 6.3, 3H), 1.09-1.16 ( , 2H), 1.25-1.40 (m, 4H), 1.92-1.97 (m, IH), 2.44 (s, 3H), 2.78-3.18 (m, 6H), 3.51 (d, 7= 6.8, 2H), 3.64 (q, 7= 6.3, IH), 3.84 (q, 7= 5.9, IH), 6.91 (d, 7= 7.0, 2H), 7.03-7.21 (m, 3H), 7.30 (d, 7= 6.8, 2H), 7.52 (d, 7= 7.1, 2H), 7.60 (d, 7= 6.9, 2H), 7.71 (d, 7= 7.1, 2H). LC-MS: 696.2 (M - H)\ 99% pure. : ' !
Example 128. Preparation of (2S,VS) 2-Λ'-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N- (7Vα-benzenesulfonyl-S-trjφtophanyl)-2,6-diaminohexanol
The title product was prepared from (2S) 2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6- diaminohexanol (example 116, step A) following the indications of general procedure Bd using Nα-benzenesulfonyl-L-tryptophan (example 4, step A). Purification by HPLC gave 93 mg (46%) of the desired material.
Η ΝMR (CDCI3X δ 0.84 (d, 7= 6.3, 3H), 0.89 (d, 7= 6.3, 3H), 0.94-1.03 (m, IH), 1.09-1.16 (m, 2H), 1.46-1.49 (m, IH), 1.93-1.98 (m, IH), 2.39 (s, 3H), 2.82-3.15 ( , 6H), 3.51 (d, 7= 6.8, 2H), 3.64 (q, = 6.3, lH),3.94 (q, 7= 5.5. IK), 6.95 (t, 7= 4.5, IH), 719 (1,7= 4.5, IH), 7.23- 7.31 (m, 6H), 7.42 (t, 7= 4.5, IH), 7.60 (d, 7= 6.8, 2H), 7.73 (d, 7 = - 6.8, 2H). LC-MS: 667.5 (M - H)\ 99% pure. ; ' Example 129. Preparation of (2S,2'S) 2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N- [7Vα-(2-thiophenesulfonyl)-S-tryptophanyI]-2,6-diaminohexanoI
The title product was prepared from (2S) 2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6- diaminohexanol (example 116, step A) following the indications of general procedure Bd using Nα-(2-thiophenesulfonyl)-L-tryptophan (example 53 , step A). Purification by HPLC gave 81 mg (40%) of the desired material.
Η ΝMR (CDC13): δ :0.84.(d, 7= 6.3, 3H), 0.89 (d, 7= 6.3, 3H), 0.94-1.03 (m, IH), 1.09-1.16 (m, 2H), 1.46-1.49 (m, IH), 1.93-1.98 (m, 1H), 2.40 (s, 3H), 2.82-3.15 (m, 6H), 3.51 (d, 7= 6.8, 2H), 3.64 (q,7= 63, 1H), 4.05 (t,7= 7.2, IH), 6.89 (1,7=4.6, lH), 6.97 (s, 1H), 7.03 (1,7= 4.5, IH), 7.19 (t, 7= 4.5, IH), 7.23-7.28 (m, 3H), 7.34-7.42 (m, 4H), 7.731 (ds 7= 6.8, 2H). LC-MS: 674.2 (M - H)-, 99% pure. '
Example 130. Preparation of (2S,2'S) 2-N-isobutyl-2-N-(4-methyIbenzenesulfonyl)-6-N- [7Vα-(4-nitrobenzenesulfonyl)-S-tryptophanyI]-2,6-diaminohexanol
The title product was prepared from (2S) 2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6- diaminohexanol (example 116, step A) following the indications of general procedure Bd using Nα-(4-nitrobenzenesulfonyl)-L-tryptophan (example 5, step A). Purification by HPLC gave 101 mg (47%) of the desired material.
T ΝMR (CDC13):(. δ 0.90-0.98 (m, 6H), 1.09-1.16 (m, 2H), 135-139 (m, 2H), 1.45-1.52 (m, IH), 1.92-1.97 (m, IH), 2.46 (s, 3H), 2.82-3.10 (m, 4H), 3.11-3.26 (m, 3H), 3.51 (d,7=7.0, 2H), 3.74 (q, 7= 6.3, IH), 3.95 (br s, IH), 6.91 (t, 7= 7.0, IH), 6.97 (s, IH), 7.01 (t, 7= 6.8, IH), 7.15 (d, 7= 6 6, IH), 7.20-7.30 ( , 3H), 7.52 (d, 7= 7.1, IH), 7.71 (d, 7= 6.9, 2H), 7.79 (d, 7 = 7.1, 2H). LC-MS: 713.1 (M - H)\ 99% pure. Example 131: Preparation of (2S,2'S) 2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N- (7Vα-acetyl-S-tryptophanyl)-2,6-diaminohexanol
The title product was prepared from (2S) 2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6- diaminohexanol (example 116, step A) following the indications of general procedure Bd using
Nα-acetyl-L-tryptophan (example 99, step A), Purification by HPLC gave 68 mg (39%) of the desired material.
Η ΝMR (CDC13): δ 0.90-0.98 (m, 6H), 1.09-1.16 (m, IH), 1.25-1.29 ( , IH), 1.45-1.52 (m, IH), 1.92-1.97 (m, IH), 2.0 (s, 3H), 2.38 (s, 3H), 2.82-3.10 (m, 4H), 3.11-3.26 (m, 3H), 3.51 (d, 7= 7.0, 2H), 3.64 (q, 7= 6.3, IH), 3.75 (br s, IH), 6.97-7.21 (nv6H), 735 (q, 7= 6.7, IH), 7.61 (d, 7= 7.1, IH), 7.69 (d, 7= 7.6, IH). LC-MS: 569.1 (M - H)\ 98% pure.
Example 132. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-[7Vα-(4- methoxyphenylacetyl)-L-phenyIaIanyI]-L-Iysine
Step A. Preparation of Nα-(4-methoxyphenylacetyl)-L-phenylalanme
L-phenylalanine was reacted with 4-methoxyphenylacetyl chloride under the conditions used in general procedure A giving the title compound which was recrystallised neat (80%) and used as such in the next step.
Step B. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-[N'α-(4- methoxyphenylacetyl)-L-phenylalanyl]-L-lysine
The title compound was prepared from Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine hydrochloride (100 mg, 0.25 mmol, example 1, step E) as described in general procedure Be. using Nα-(4-methoxyphenylacetyl)-L-phenylalanine (80 mg, 0.25 mmol) which was prepared in step A of this example. The final product was triturated with ether to yield 85 mg (18%) of the desired material. Η NMR (CDC13): δ 0.83 (d, 7= 6.9, 6H), 1.08-1.11 (m, 2H), 133-1.55 (m, 2H), 1.45-1.52 (m, IH), 1.79-1.89 (m, 2H), 2.36 (s, 3H), 2.85- 3.27 (m, 6H), 3.55 (s, 2H), 3.79 (s, 3H), 4.21 (s, 2H), 4.53 (t,7= 5.9, IH), 6.79 (d,7= 8.2, 2H), 6.99-7.09 (m, 3H), 7.15-7.26 (m, 7H), 7.73 (d,7= 8.1, 2H). LC-MS: 650.1 (M - H)\ 98% pure.
Example 133. Preparation of Nα-isoburyl-Nα-(4-methylbenzenesulfonyl)-Ne-(7Vα- dihydrocinnamoyl-L-tryptophanyl)-L-lysine
Step A. Preparation of Nα-dihydrocirmamoyl-L-tryptophan
L-tryptophan was reacted with Nα-dihydrocinnamoyl chloride under the conditions used in general procedure A giving the title compound which was recrystallised from DCM (55%). This material was used without further purification in the next step.
Step B. Preparation ofΛ?α-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-(N -dihydrocinnamoyl- L-tryptophanyl)-L-Iysine
The title compound was prepared from Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine hydrochloride (400 mg, 1.0 mmol, example 1 , step E) as described in general procedure Be using Nα-dihydrocinnamoyl-L-tryptophan (360 mg, 1.0 mmol) which was prepared in step A of this example. The final product was triturated with ether to yield 612 mg (85% pure) of the crude material. Purification of 100 mg of the crude material by HPLC gave 45 mg (45%) of pure adduct.
]H ΝMR (CDC13): δ 0.82 (d, 7= 6.8, 3H), 0.84 (d, 7= 6.8, 3H), 1.08-1.11 (m, 2H), 1.18-1.22 (m, 2H), 1.45-1.52 (m, IH), 1.94-1.99 (m, 2H), 2.34 (s, 3H), 2.48-2.52 (m, 2H), 2.89-3.09 (m, 4H), 3.15-3.18 (m, 2H), 4.29 (br s, IH ), 4.77 (br s, IH), 6.95 (t, 7= 7.2, IH), 7.04-734 (m, 8H), 736 (d, 7= 8.1, IH), 7.69 (d, 7= 7.2, IH), 7.99 (d, 7= 8.1, 2H). LC-MS: 673.1 (M - H)\ 98% pure. Example 134. Preparation of (2S,TS) 2-N-isobutyl-2-N-(4-methylbenzenesulfonyI)-6-N- (7Vα-te?r/-butoxycarbonyI-S-2-ρyridyIalanyI)-2,6-diaminohexanol
The title product was prepared from (2S) 2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6- diaminohexanol (example 116, step A) following the indications of general procedure Bd using Nα-tert-butoxycarbonyl-L-2-pyridylalanine (Peptech Corporation). Purification by HPLC gave 54 mg (30%)) of the desired material.
'H ΝMR (CDC13): δ 0.90-0.98 (m, 6H), 1.09-1.16 (m, IH), 1.25-1.29 (m, 2H), 1.45 (s, 9H), 1.92-1.97 (m, IH), 2.38 (s, 3H), 2.82-3.10 (m, 4H), 3.26 (br s, 2H), 3.51 (d, 7= 7.0, 2H), 3.64 (q, 7= 6.3, IH), 4.55 (br s, IH), 7.18-7.38 (m, 4H), 7.70 (t, 7= 6.8, IH), 7.74 (d, 7=J.5, 2H), 8.45 (s, IH). LC-MS: 591.3 (M + H)+, 98% pure.
Example 135. Preparation of (2S,TS) 2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N- (7Vα-tert-butoxycarbonyI-S-3-pyridyIaIanyI)-2,6-diaminohexanol
The title product was prepared from (25) 2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6- dia inohexanol (example 116, step A) following the indications of general procedure Bd using Nα-tert-butoxycarbonyl-L-3-pyridylalanine (Peptech Corporation). Purification by HPLC gave 45 mg (25%) of the desired material.
'H MR (CDC13): δ 0.90-0.98 (m, 6H), 1.09-1.16 (m, IH), 1.25-1.29 (m, 2H), 1.45 (s, 9H), 1.92-1.97 (m, IH), 2.37 (s, 3H), 2.82-330 (m, 6H), 3.51 (d, 7= 7.0, 2H), 3.64 (q, 7= 6.3, IH), 435 (br s, IH), 7.18-7.38 (m, 4H), 7.60 (br s, IH), 7.74 (d, 7= 7.5, 2H), 8.56 (d, 7= 6.6, 2H). LC-MS: 591.3 (M + H)+, 95% pure.
Example 136. Preparation of (2S,2'S) 2-N-isobutyI-2-Λ;-(4-methylbenzenesulfonyl)-6-N- (7Vα-tert-butoxycarbonyl-S-4-pyridyIaIanyI)-2,6-diaminohexanol
The title product was prepared from (25) 2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6- diaminohexanol (example 116, step A) following the indications of general procedure Bd using Nα-/cτ/-buloxycarbony]-L-4-pyridy]alanine (Peptech Corporation). Purification by HPLC gave 65 mg (36%>) of the desired material.
'H ΝMR (CDC13): δ 0.90-0.98 (m, 6H), 1.09-1.16 (m, IH), 1.25-1.29 (m, 2H), 1.49 (s, 9H), 1.92-1.97 ( , IH), 237 (s, 3H), 2.82-330 (m, 6H), 3.49-3.51 (m, 2H), 3.64 (br s, IH), 4.35 (br s, IH), 7.16 (s, 2H), 7.18-7.22 (m, 2H), 7.74 (d, 7= 7.5, 2H), 8.56 (br s, 2H). LC-MS: 591.3 (M + H)+, 98% pure.
Example 137. Preparation of (2S,2'S) 2-N-isobutyl-2-N-(4-methyIbenzenesuIfonyI)-6-N- (7Vα-tert-butoxycarbonyl-S-4-thiazolylaIanyl)-2,6-diaminohexanol
The title product was prepared from (2S) 2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6- diaminohexanol (example 116, step A) following the indications of general procedure Bd using Nα-tert-butoxycarbonyl-L-4-thiazolylalanine (Peptech Corporation). Purification by HPLC gave 50 mg (29%) of the desired material.
'H ΝMR (CDC13): δ 0.90-0.98 (m, 6H), 1.09-1.16 (m, IH), 1.25-1.29 (m, 2H), 1.49 (s, 9H), 1.92-1.97 (m, IH), 2.37 (s, 3H), 2.82-330 (m, 6H), 3.49-3.51 (m, 2H), 3.64 (br s, IH), 4.35 (br s, 3 H), 7.06 (s, IH), 7.18-7.22 (m, 3H), 7.74 (d,7= 7.5, 2H). LC-MS: 697.3 (M + H)+, 99% pure.
Example 138. Preparation of (2S,2'S) 2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N- (7Vα-ter/-butoxycarbonyl-S-2-fluorophenylalanyl)-2,6-diaminohexanol
The title product was prepared from (25) 2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6- diaminohexanol (example 116, step A) following the indications of general procedure Bd using Nα-te7't-butoxycarbonyl-L-2-fluorophenylalanine (Peptech Corporation). Purification by HPLC gave 62 mg (34%) of the desired material.
Η ΝMR (CDC13): δ 0.79 (d, 7= 63, 3H), 0.82 (d, 7= 6.3, 3H), 1.08-1.11 (m, 2H), 1.29-134 (m, 3H), 1.41 (s, 9H), 1.92-1.97 (m, IH), 2.38 (s, 3H), 2.84-3.09 (m, 6H), 3.51 (d, 7= 7.1, 2H), 3.72 (t, 7= 6.1, IH), 4.35 (br s, IH), 7.00-7.08 (m, 2H), 7.14-7.28 (m, 4H), 7.73 (d,7= 7.9, 2H). LC-MS: 608.4 (M + H)+, 99% pure.
Example 139. Preparation of (2S,2'S) 2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N- (7Vα-tert-butoxycarbonyl-S-3-fluorophenylaIanyI)-2,6-diaminohexanoI
The title product was prepared from (25) 2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6- diaminohexanol (example 116, step A) following the indications of general procedure Bd using Nα-tert-butoxycarbonyl-L-3-fluorophenylalanine (Peptech Corporation). Purification by HPLC gave 88 mg (48%) of the desired material. • •
Η ΝMR (CDC13): δ 0.85 (d, 7= 6.3, 3H); 0.90 (d, 7= 6.3, 3H), 1.08-1.11 (m, 2H), 1.29-134 (m, 3H), 1.41 (s, 9H), 1.92-1.97 (m, 1H), 2.41 (s, 3H), 2.84-3.09 (m, 6H), 3.51 (d, 7= 7.1, 2H), 3.62 (t, 7= 6.1, IH), 4.35 (br s, IH), 6.91 (d, 7= 6.6, 2H), 7.00 (d, 7= 6.7, 2H), 7.14-7.28 (m,
3H), 7.73 (d, 7= 7.9, 2H). LC-MS: 608.4 (M + H)+, 99% pure.
Example 140. ' Preparation of (2S,TS) 2-N-isoburyl-2-N-(4-methylbenzenesulfonyl)-6-N- (7Vα-tert-butoxycarbonyl-S-4-fIuorophe ylalanyl)-2,6-diaminohexanol
The title product was prepared from (2S) 2-N-isobutyl-2-N-(4-m,ethylbenzenesulfonyl)-2,6- diaminohexanol (example 116, step A) following the indications of general procedure Bd using Nα-tert-butoxycarbonyl-L-4-fluorophenylalanine (Peptech Corporation). Purification by HPLC gave 65 mg (35%) of the desired material.
Η ΝMR (CDC13): δ 0.85 (d, 7= 6.3, 3H), 0.90 (d, 7= 6.3, 3H), 1.08-1.11 (m, 2H), 1.29-1.34 (m, 3H), 1.41!(s, 9H), 1.92-1.97 (m, IH), 2.41 (s, 3H), 2.84-3.09 (m, 6H), 3.51 (d, 7= 7.1, 2H), 3.60-3.62 (m,1 IH), 4.24 (br s, IH), 6.94 (t, 7= 7.6, 2H), 7.14-7.18 (ni, 2H), 7.20-730 (m, 2H),
7.73 (d, 7= 7.9, 2H). LC-MS: 608.4 (M + H)\ 99% pure. , Example 141. Preparation of (2S,2'S) 2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N- (Λr'α-t'ert-butoxycarbonyl-S-l,2,3,4-tetrahydronorharman-3-carbonyl)-2,6- diaminohexanol
The title product was prepared from (25) 2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6- diaminohexariol (example 116, step A) following the indications of general procedure Bd using Nα-tert-butoxycarbonyl-L- 1 ,2,3 ,4-tetrahydronorharman-3-carboxylic acid (Peptech Corporation). Purification by HPLC gave 75 mg (39%) of the desired material.
Η ΝλfR (CDCl3): δ 0.85 (d, 7= 6.3, 3H), 0.90 (d, 7= 6.3, 3H), 1.08-1.11 (m, 2H), 1.29-134 (m, 3H), 136:(s, 9H), 1.92-1.97 (m, IH), 2.35 (s, 3H), 2.40 (t, 7= 6.7, IH), 2.80-3.09 (m, 6H), 3.52 (d, 7= 5.1, 2H), 3.60-3.62 ( , IH), 4.23 (br s, IH), 7.28 (d, 7= 7.8, 2H), 7.73 (d, 7= 7.9, 2H). LC-MS: 641.5 (M + H)+, 99% pure. " ' .
Example 142. Preparation of (2S,TS) 2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N- (TV α-tert-butoxy carbonyl-S-4-tert-butylphenylalanyl)-2,6-diaminohexanol
The title product was prepared from (2S) 2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6- diaminohexanol (example 116, step A) following the indications of general procedure Bd using Nα-te7't-butoxycarbonyl-L-4-tert-butylphenylalanine (Peptech Corporation). Purification by HPLC gave 81'irig (42%) of the desired material.
' , i ' . Η ΝMR (CDCi3): δ 0.85 (d, 7= 6.3, 3H), 0.90 (d, 7= 6.3, 3H), 1.08-1.11 (m, 2H), 1.29-134
(m, 3H), 1.36 (s, 9H), 1.92-1.97 (m, IH), 2.40 (s, 3H), 2.80-3.29 (m, 8H), 3.52 (d, 7= 5.1, 2H),
3.60-3.62 (m, IH), 4.23 (brs, IH), 6;90-7.28 (m, 5H), 7.55 (d,7=6.6, IH)), 7.73 (d,7= 7.9, 2H).
LC-MS: 646÷5 (M + H)+, 99% pure. Example 143. Preparation of (2S,2'S) 2-N-isobutyI-2-N-(4-methylbenzenesuIfonyl)-6-N- (/Vα-tert-butoxycarbonyl-S-pentafIuorophenyIalanyl)-2,6-diaminobexanol
The title product was prepared from (25) 2-N-isobutyI-2-N-(4-methyIbenzenesulfonyl)-2,6- diaminohexanol (example 116, step A) following the indications of general procedure Bd using Nα-tert-butoxycarbonyl-L-pentafluorophenylalanine (Peptech Corporation). Purification by HPLC gave 55 mg (27%) of the desired material.
'H ΝMR (CDC13): δ 0.85 (d, 7= 6.3, 3H), 0.90 (d57= 6.3, 3H), 1.08-1.11 ( , 2H), 1.29-134 (m, 3H), 1.41 (s, 9H), 1.92-1.97 (m, IH), 2.41 (s, 3H), 2.80-3.09 (m, 6H), 3.51 (d, 7= 7.1, 2H), 3.60-3.62 (m, IH), 4.31 (br s, IH), 7.09 (s, 2H ), 7.10-7.28 (m, 5H), 7.68 (d, 7= 7.5, 2H). LC- MS: 680.2 (M + H)+, 99% pure.
Example 144. Preparation of (2S,2'S) 2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N- [7Vα-tert-butoxycarbonyl-S-4-(9-fluorenemethoxycarbonylaminomethyl) phenylaIanyI]-2,6-diammohexanoI
The title product was prepared from (25) 2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6- diaminohexanol (example 116, step A) following the indications of general procedure Bd using Λrα-ter/-butoxycarbonyl-L-4-(9-fluorenemethoxycarbonylaminomethyl) phenylalanine (Peptech Corporation). Purification by HPLC gave 41 mg (16%) of the desired material.
LC-MS: 842.8 (M + H)+, 99% pure.
Example 145. Preparation of (2S,2'S) 2-N-(4-aminobenzenesulfonyl)-2-Λ-isobutyl-6-N- [7V,α-(4-methylbenzenesulfonyl)-S-tryptophanyl]-2,6-diaminohexanol
Step A. Preparation of Nα-isobutyl-Λα-(4-nitrobenzenesulfonyl)-Ne-benzyloxycarbonyl-L-lysine methyl ester
This product was prepared following the procedure described for the preparation of Nα-isobutyl- Nα-(4-methylbenzenesuIfonyl)-Ne-benzyloxycarbonyl-L-lysine methyl ester (example 65, step B) using 4-nitrobenzenesulfonyl chloride instead of 4-methylbenzenesulfonyl chloride. The yield of this reaction was 42%.
Η ΝMR (CDC13): δ 0.83 (d, 7 = 6.6, 3H), 0.86 (d, 7 = 6.5, 3H), 135-1.69 (m, 5H), 1.88-2.00 (m, 2H), 2.90 and 3.04 (ABX,7= 14.5, 7.5, 2H), 3.18 (m, 2H), 3.49 (s, 3H), 4.45 (t,7 = 6.0, IH), 4.83 (s, IH), 5.10 (s, 2H), 7.30-7.40 (m, 5H), 8.00 (d, 7 = 8.5, 2H), 8.33 (d, 7 = 8.5, 2H).
Step B. Preparation of Nα-(4-aminobenzenesulfonyl)-Nα-isobutyl-L-lysine methyl ester
The title compound was obtained by catalytic hydrogenation of Nα-isobutyl-Nα-(4- nitrobenzenesulfonyl)-Ne-benzyloxycarbonyl-L-lysine methyl ester (step A) as described for the preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine methyl ester (example 65, step C). This compound was used without purification in the next step.
Step C. Preparation of Nα-(4-aminobenzenesulfonyl)-Nα-isobutyl-Ne-[N'α-(4- methylbenzenesulfonyl)-L-tryptophanyl]-L-lysme methyl ester
The title compound was prepared in the same manner than example 69 using Nα-(4- arninobenzenesulfonyl)-Niα-isobuty]-L -lysine methyl ester (step B) as the starting material. The yield of this reaction was 65%.
Η ΝMR (DMSO-d6): δ 0.77 (t, 7 = 7.5, 6H), 0.90-1.10 (m, 4H), 1.40 (m, IH), 1.70 ( , IH), 1.85 (m, IH), 2.29 (s, 3H), 2.87-3.00 (m, 3H), 3.44 (s, 3H), 3.85 (m, IH), 4.10 (s, 2H), 4.24 (1, 7 = 7.3, IH), 5.74 (s, 2H), 6.60 (d, 7 = 8.4, 2H), 6.90 (m, IH), 7.00 (m, 2H), 7.15 (d, 7 = 7.5, 2H), 7.30-7.45 (m, 6H), 770 (t, 7 = 5.0, IH), 7.82 (d, 7 = 8.7, IH), 10.70 (s, IH).
Step D. Preparation of (25,2'S) 2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-6-N-[Λpα-(4- methylbenzenesulfonyl)-5-tryptophanyl]-2,6-diaminohexanol Nα-(4-am oberizenesulfonyl)-Nα-isobutyl-Ne-[Nα-(4^
L-lysine methyl ester (step C) was reduced with LiAlH4 following the indications of general procedure G. The final product was obtained in 37% yield.
'HΝMR (DMSO-d6): δ 0.80 (d, 7= 7.8, 3H), 0.83 (d, 7= 7.6, 3H), 0.88-1.22 (m, 5H), 1.48 (m, IH), 1.85 (m, IH), 2.30 (s, 3H), 2.65-2.80 (m, 4H), 2.85 (dd, 7= 14.5, 7.5, IH), 2.93 (dd, 7 = 143, 7.6, IH), : 3.20-3.40 (m, 2H), 3.45 (m, l'H), 3.88 (m, IH), 4.60 (t, 7= 5.0, lH), 5.90 (s, 2H), 6.60 (d, 7= 8.4, 2H), 6.90 ( , IH), 7.00 ( , 2H), 7.15 (d, 7= 7.5, 2H), 730-7.45 (m, 6H), 7.70 (1, 7= 5.0, IH), 7.82 (d, 7= 8.7, IH), 10.70 (s,'lH).
Example 146. Preparation of (2S,2'S) 2-N-isobutyl-2-N-(4-methoxybenzenesulfonyl)-6-N- (S-tryptophanyl)-2,6-diaminohexanol trifluoroacetic acid salt
This product was obtained quantitatively by treating (25,2'5) 2-N-isobutyl-2-N-(4- ■ methoxybenzenesulfonyl)-6-N-(N,α-tert-butoxycarbonyl-5-tryptophanyl)-2,6-diaminohexanol (example 147) with TFA in CH2C12.
Η ΝMR (DMSO-d6): δ 0.80 (d, 7= 7.0, 3H), 0.83 (d, 7= 7.0, 3H), 0.92-1.60 (m, 6H), 1.88 (m, IH), 2.78-3.00 (m, 4H), 3.05 and 3.20 (ABX, 7= 14.0, 7.0, 2H), 3.22 and 3.30 (ABX, 7= 14.2, 7.0, 2H), 3.50 (m, IH), 3.80 (s, 3H), 3.90 (m, IH), 4.35 (m, lH), 6.95-7.12 (m, 4H), 7.18 (s, IH), 7.38 (d, 7= 8.2, IH), 7.60 (m, IH), 7.70 (d, 7= 8.2, 2H), 8.05 (brs, 3H), 834 (m,'lH), 11.0 (s, rn).;, , ; x ' j ' '
I . -
Example 147. Preparation of (2S,TS) 2-N-isobutyl-2-N-(4-methoxybenzenesulfonyl)-6-N- (Λ^α-^rt-butoxycarbonyl-S-tryptophany^^.θ-diammohexanol
Step A. Preparation of Nα-isobutyl-Nα-(4-methoxybenzenesulfonyl)-N6-benzyloxycarbonyl-L- lysine methyl ester
This product was prepared following the procedure described for the preparation of Nα-isobutyl- Nα-(4-methylbenzenesulfonyl)-Ne-benzyloxycarbonyl-L-lysine methyl ester (example 65, step B) using 4-methoxybenzenesulfonyl chloride instead of 4-methylbenzenesulfonyl chloride. The yield of this reaction was 65%.
'H NMR (CDCI3): δ 0.84 (d, 7 = 6.6, 3H), 0.85 (d, 7 = 6.5, 3H), 130-1.65 (m, 5H), 1.85-1.95 (m, 2H), 2.90 and 3.04 (ABX, 7 = 14.5, 7.5, 2H), 3.15 (m, 2H), 3.51 (s, 3H), 3.85 (s, 3H), 4.35 (t, 7= 5.5, IH), 4.80 (br s, IH), 5.09 (s, 2H), 6.94 (d, 7 = 8.6, 2H), 730-7.40 (m, 5H), 7.25 (d, 7 = 8.5, 2H). ' ., • ',
Step B. Preparation of Nα-isobutyl-Nα-(4-methoxybenzenesulfonyl)-L-lysine methyl ester
The title compound was obtained quantitatively by catalytic hy drogenation of Nα-isobutyl-Nα-(4- methoxybenzenesulfonyl)-Ne-benzyloxycarbonyI-L-lysine methyl ester (step A) as described for the preparation of Λα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine methyl ester (example 65, step C). This compound was used without purification in the next step.
Step C. Preparation . of Nα-isobutyLNα-(4-methoxybenzenesulfonyl)-Ne-(N'α-tert- butoxycarbonyl-L-tryptophanyl)-L-lysine methyl ester
The title compound1 was prepared in the same manner" as in the example 65 (step D) using Nα- isobutyl-Nα-(4-methoxybenzenesulfonyl)-L-lysine methyl ester (step B) and commercially available Nα-te7Y-butoxycarbonyl-L-tryptophan as the starting materials. The yield of this reaction was 86%.
Η ΝMR (DMSO-d6): δ 0.84 (t, 7 = 7.0, 6H), 0.90-1.30 (m, 5H), 1.31 (s, 9H), 1.53 (m, IH), 1.86 (m, IH), 2.82-3.10!(m, 5H), 3.46 (s, 3H), 3.83 (s, 3H), 4.12 (m, IH), 4.30 (t, 7 = 5.0, IH), 6.67 (d, 7 = 8.2, IH), 6.90-7.12 (m, 5H), 7.30 (d, 7= 8.0, 2H), 7.55 (m, IH), 7.70 (m, 2H), 7.81 (m, iH). . ■; :■ ■' ■ ' : ■ ' . - " _
)
Step D. Preparation of (25,2'S) 2-N-isobutyl-2-N-(4-methoxybenzenesulfonyl)-6-N-(N'α-tgrt- butoxycarbonyl-S-tryptophanyl)-2,6-diaminohexanol Nα-isobutyl-Nα-(4-methoxybenzenesulfonyl)-Ne-(N,α-tert-butoxycarbonyl-L-tryptophanyl)-L- lysine methyl ester (step C) was reduced with LiAlH4 following the indications, of general procedure G. The final product was obtained in 81% yield.
'H ΝMR (DMSO-d6): δ 0.73 (t,7=7.0, 6H), 0.90-1.27 (m, 5H), 1.31 (s, 9H), 1.52 (m, IH), 1.85 (m, IH), 2.80 and 3.02 (ABX, 7= 14.0, 7.2, 2H), 2.90 (m, 2H), 3.50 (m, IH), 3.82 (s, 3H), 4.15 (m, IH), 4.65 (t,7= 5.0, IH), 6.65 (d,7= 7.8, IH), 6.92-7.12 (m, 5H), 7.30 (d,7= 7.8, IH), 7.58 (d, 7= 8.0, IH), 7.70 (d, 7= 7.7, 2H), 7.73 (t, 7= 5.0, IH), 10.77 (s, IH).
Example 148. Preparation of (2S,2'S) 2-N-isobutyl-2-N-(4-methoxybenzenesulfonyl)-6-N- (/Vα-pϊvaloyl-S-tryptophanyl)-2,6-diaminohexanol
Treatment of (2S,2'5)2-N-isobutyl-2-N-(4-methoxybenzenesulfonyl)-6-N-(S-tryptophanyl)-2,6- diaminohexanol trifluoroacetic acid salt (example 146) with pivaloyl chloride using the conditions described in example 65 (step B) afforded the desired product in 86% yield!
'H ΝMR (DMSO-d6): δ.0.80 (d, 7= 7.0, 3H), 0.83 (d, 7= 7.0, 3H), 1.02 (s, 9H), 1.03-1.30' (m, 5H), 1.53 (m, IH), 1.88 (m, IH), 2.80 (dd, 7= 14.5, 7.5, IH), 2.83-3.10 (m, 5H), 3.27 and 3.32 (ABX, 7= 14.5, 7.2, 2H), 3.52 (m, IH), 3.82 (s, 3H), 4.46 (m, IH), 6.90-7.15 (m, 5H), 7.30 (d, 7= 8.1, IH), 7.56 (d, 7= 8.0, IH), 7.71 (d, 7= 8.2, 2H), 7.74 (t, 7= 5.0, IH), 10.75 (s, IH).
Example 149. Preparation of (2S,2'S) 2-N-isobutyl-2-N-(4-methoxybenzenesulfonyl)-6-N- [7Vα-(4-methylbenzenesulfonyl)-S-tryptophanyl]-2,6-diammohexanol
Treatment of (25,2'S)2-N-isobutyl-2-N-(4-methoxybenzenesulfonyl)-6-Λ''-(5-tryptophanyl)-2,6- diaminohexanol trifluoroacetic acid salt (example 146) with 4-methylbenzenesulfonyl chloride using the conditions described in example 65 (step B) afforded the desired product in 82% yield.
Η ΝMR (DMSO-d6): δ 0.80 (d, 7= 6.0, 3H), 0.83 (d, 7= 6.2, 3H), 0.88-1.50 (m, 6H), 1.88 (m, IH), 2.30 (s, 3H), 2.65 (m, 2H), 2.78 (m, 2H), 2.93 (m, 2H), 3.22-3.40 (m, 2H), 3.50 (m, IH), 3.82 (s, 3H), 3.86 (m, IH), 4.67 (m, IH), 6.90 (t,7= 7.5, IH), 7.00-7.18 (m, 6H), 7.28 (d,7= 83, lH), 734 (d,7= 8.0, lH), 7.45 (d,7= 7.5, 2H), 7Jl (m, 3H), 7.83 (d,7=7.4, 1H), 10J1 (s, IH).
Example 150. Preparation of Nα-cyclopentylmethyl-Nα-(4-methoxybenzenesulfonyl)-Ne- (7Vα- 'ert-butoxycarbonyl-L-tryptophanyl)-L-lysine methyl ester
Step A. Preparation of Nα-cyclopentylmethyl-Nα-(4-methόxybenzenesulfonyl)-Ne- benzyloxycarbonyl-L-lysine methyl ester
This product was prepared following the procedure described for the preparation of Nα-isobutyl- Nα-(4-methylbenzenesulfonyl)-Ne-benzyloxycarbonyl-L-lysine methyl ester (example 65, steps A and B) using cyclopentanecarboxaldehyde instead of isobutyraldehyde. (for step A) and 4- methoxybenzenesulfonyl chloride instead of 4-methylbenzenesulfonyl chloride (for step B). The yield of this two-step sequence was 79%.
ΗΝMR (CDC13): δ 1.03-1.45 (m, 6H), 1.48-1.71 (m, 8H), 1.56 (s, 9H), 1.92 (m, 1H), 2.18 (m, IH), 3.03 and 3.13 (ABX, 7= 13.5, 7.0, 2H), 3.20 (m, 2H), 3.53 (s, 3H), 3.87 (s, 3.H), 4.40 (t, 7 = 6.5, IH), 5.11 (s, 2H),' 6.96 (d, 7= 8.7, 2H), 7.25-7.40 (m, 6H), 7.77 (d, 7= 8.7, 2H).
Step B. Preparation of Nα-cyclopentylmethyl-Nα-(4-methoxybenzenesulfonyl)-L-lysine methyl ester ' '
The title compound was obtained quantitatively by catalytic hydrogenation of Nα- cyclopentylmethyl-Nα-(4-memoxybenzenesulfonyl)-N€-berιzyloxycaι-bonyl-L-lysine methyl ester as described for the preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine methyl ester (example 65, step C). This compound was used without purification in the next step.
Step C. Preparation of Nα-cyclopentylmethyl-Nα-(4-methoxybenzenesulfonyl)-Ne-(N'α-tgrt-
• • • i butoxycaτbonyl-L-tryptophanyl)-L-lysine methyl ester
The title compound was prepared in the same manner as in example 65 (step D) using Nα- cyclopentylmethyl-Nα-(4-methoxybenzenesulfonyl)-L-lysine methyl ester (this example, step B) andNα-tert-butoxycarbonyl-L-tryptophan-N-hydroxysuccinimide as the starting materials. The yield of the two last reactions was 82%).
'HΝMR (CDC13): δ 1.08-1.28 (m, 6H), 1.30 (s, 9H), 1.40-1.62 (m, 7H), 1.80 (m, IH), 2.10 (m, IH), 2.82-3.10 (m, 5H), 3.46 (s, 3H), 3.82 (s, 3H), 4.12 (m, IH), 430 (m, IH), 6.65 (d, 7= 8.2, IH), 6.90-7.10 (m, 5H), 7.30 (d, 7= 8.0, IH), 7.59 (d, 7= 8.2, IH), 770 (d, 7= 8.1, 2H), 7.80 (t, 7= 5.0, IH), 10.77 (s, IH).
Example 151. Preparation of Nα-cyclopentylmethyl-Nα-(4-methoxybenzenesulfonyl)-Ne- (TVα-tert-butoxycarbonyl-L-tryptophanyl)-L-lysiπe
A saponification of Nα-cyclopentylmethyl-Nα-(4-methoxybenzenesulfonyl)-Ne-(N,α-tert- butoxycarbonyl-L-tryptophanyl)-L-lysine methyl ester (example 150) using the conditions described in example 65 (step E) afforded the desired material quantitatively.
>HΝMR (DMSO-d6): δ 1.08-1.28 (m, 6H), 1.21 (s, 9H), 1.40-1.65 (m, 7H), 1.80 (m, IH), 2.18 (m, IH), 2.88-3.10 (m,'6H), 3.82 (s, 3H), 4,12 (m, IH), 4.20 (m, IH), 6.62 (d,7= 7.8, IH), 6.90- 7.15 (m, 5H), 7.3 (d, 7= 8.0, IH), 7.55 (d, 7= 8.0, IH), 7.70 (d, 7= 8.7, 2H), 7.80 (d, 7= 5.1, IH), 10.77 (s, IH), 12.70 (br s, IH).
Example 152. Preparation of (2S,2'S) 2-N-cyclopentylmethyl-2-N-(4- methoxybenzenesulfonyl)-6-N-(S-tryptophanyl)-2,6-diaminohexanol trifluoroacetic acid salt
This product was obtained quantitatively by treating (25,2'5) 2-N-cyclopentylmethyl-2-N-(4- methoxybenzenesulfonyl)-6-N-(Nα-tert-butoxy carbonyl- S-tryptophanyl)-2,6-diaminohexanol (example. 153) with TFA in CH2C12. ΗNMR (DMSO-d6): δ 1.10-138 (m, 7H), 1.40-1.68 (m, 7H), 2.18 (m, IH), 2.84-3.02 (m, 4H), 3.10 and 3.18 (ABX, 7= 14.0, 7.0, 2H), 3.30 (m, 2H), 3.50 ( , IH), 3.81 (s, 3H), 3.90 (m, IH), 4.35 (m, IH), 6.98-7.10 (m, 4H), 7.20 (s, IH), 7.39 (d, 7= 83, IH), 7.60 (m, IH), 7.70 (d, 7= 8.2, 2H), 8.10 (br s, 3H), 8.36 (m, IH), 11.0 (s, IH).
Example 153. Preparation of (2S,2'S) 2-N-cyclopentylmethyl-2-N-(4- methoxyberιzenesulfonyl)-6-N-(Λ7,α-fert-butoxycarbonyl- S-tryptophanyl)- 2,6-diaminohexanol
Nα-cyclopentylmethyl-Nα-(4-methoxybenzenesulfonyl)-Ne-(N'α-tert-butoxycarbonyl-L- tryptophanyl)-L-lysine methyl ester (example 150) was reduced with LiAlH4 following the indications of general procedure G. The final product was obtained With 72%) yield.
ΗΝMR (DMSO-d6): δ 0.95-1.28 (m, 7H), 1.30 (s, 9H), 1.42-1.65 (m, 6H), 2.15 (m, IH), 2.85- 3.10 (m, 6H), 3.35 (m, IH), 3.53 (m, IH), 3.80 (s, 3H), 4.15 (m, IH), 4.65 (m, IH), 6J5 (d, 7= 8.2, IH), 6.90-7.12 (m, 5H), 7.30 (d, 7= 8.0, IH), 7.55 (d, = 8.4, IH), 7.70 (d, 7= 8.0, 2H), 7.73
(t, 7 = 5.0, IH). ' ; ■ ' ! . i ■ . ' ■ .
Example 154. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-[7V,α-(4- m ethy lb enzenesulf ony l)-L-ph eny lalanyl] -L-o rnithin e
The title compound was prepared using the conditions described in example 65 (steps A, B, C, D and E) With, as the starting materials, . Ne-benzyloxycarbonyl-L-ornithine methyl ester hydrochloride and, for step D, Nα-(4-methylbenzenesulfonyl)-L-phenylalanine.
Η ΝMR (DMSO-d6): δ 0.77 (d, 7= 6.5, 3H), 0.80 (d, 7= 6.8, 3H), 1.15-1.25 (m, 2H), 1.40 (m, IH), 1.70 (m, IH), 1.90 (m, IH), 2.33 (s, 3H), 2.36 (s, 3H), 2.60-3.00 (m, 5H), 3.85 (m, 2H), 4.19 (m, 2H), 4.19 (m, IH), 7.08-7.26 (m, 6H), 7.38 (d, 7= 8.2, 2H), 7.45 (m, 2H), 7.70 (d, 7= 83, 2H), 7.35 (t, 7= 5.0, IH), 7.90 (d, 7= 8.5, IH), 8.16 (d, 7= 8.6, IH), 12.70 (br s, IH). Example 155. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-(N'α7V,α- dib enzylgly cy!)-L-lysine
The title compound was prepared from Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-(N'α- benzylglycyl)-L-lysine (100 mg, 0.20 mmol, example 109) by following the indications of general procedure H using Et3Ν (70 μL, 0.50 mmol) and benzyl bromide (0.10 mL, 0.84 mmol). The crude material ,was purified by preparative HPLC. The product was isolated as a solid (21 mg, 18% yield).
Η NMR.(CDC13): δ 0.78 (d, 7= 6.7, 3H), 0,83 (d, 7= 6.8, 3H), 134 (m, 2H), 1.45 (m, IH), 1.65 (m, 2H), 1.91 (m, 2H), 2.39 (s, 3H), 2.86 (dd, 7= 13.6, 73, lHa), 2.98 (dd, 7= 13.6, 7.3, ■lHb), 3.12 (s, IH), 3.15 (m, 2H), 3.64 (s, 4H), 4.43 (t, 7= 7.0, IH), 4.93 (s, 2H), 7.13 (s, IH), 7.16 (d, 7= 7.7, 2H), 7.22-737 (m, 10H), 7.66 (d,7= 7.6, 2H). LC-MS: 592 (M - H)\ 99% pure.
Example 156. Preparation of (2S) 2-Λr-isobutyI-2-N-(4-methylbenzenesulfonyl)-6'-N-(N'α- benzyl-NVphenylglycyl)-2,6-diaminohexanol
The title product was prepared from (25) 2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6- diaminohexanol (350 mg, 1.0 mmol, example 116, step A) following the indications of general procedure Ba using N-benzyl-N-phenylglycine (241 mg, 1.0 mmol, example 114, step A) and NN-carbonyldiimidazole (195 mg, 1.2 mmol). The crude material was purified by preparative HPLC- The product, was isolated as a solid (135 mg, 24% yield).
]HΝMR (DMSO-d6): δ 0.83 (t,7= 6.8, 6H), 1.05 (m, 2H), 1.26 (m, 3H), 1.55 (m, IH), 1.85 (m, IH), 2.37 (s, 3H), 2.86 (m, IH), 2.94 (m, 3H), 3.27 (m, IH), 3.45 (m, IH), 3.94 (s, 2H), 4.62 (s, 2H), 4.66 (t, 7= 5.1, IH), 6.57 (d, 7= 8.3, 2H), 6.61 (t, 7= 7.5, IH), 7.11 (t, 7= 7.5, 2H), 7.25 (m, 3 H), 7.31 (t, 7= 7.4, 2H), 7.35 (d, 7= 7.8, 2H), 7.66 (d, 7= 8.2, 2H), 8.81 (t, 7 - 5.0, IH). LC-MS: 566 (M + H)+, 99% pure. Example 157. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-(N,α- cyclohexylglycyl)-L-lysine
The title compound was prepared from Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne- iodoacetyl-L-lysine (520 mg, 1.0 mmol, example 105, step B) by following the indications of general procedure H using cyclohexylamine (1.0 mL, 8J mmol). The crude material was purified by preparative HPLC. The product was isolated as a solid (300 mg, 64%> yield). .
Η ΝMR (CDCLTMeOD 1 :1): δ 0.85 (d, 7= 7.2, 3H), 0.88 (d, 7= 5.8, 3H), 1.23 (m, IH,), 130-1.50 (m, 6H), 1.56 (m, 3H), 1.70 (m, IH), 1.88 (m, 3H), 2.04 (m, 3H), 2.41 (s, 3H), 2.98 (m, IH), 3.07 (m, 2H), 3.26 (t, 7= 6.4, 2H), 3.66 (s, 2H), 4.27 (t, 7= 7.3, IH), 7.31. (d, 7= 8.4, 2H), 7.75 (d, 7= 8.8, 2H). LC-MS: 494 (M - H)", 95% pure.
Example 158. Preparation of (2S) 2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N-(N'α- benzoyl-N'α-benzylglycyl)-2,6-diaminohexanol
Step A. Preparation of N-benzylglycine methyl ester
A solution of methyl bromoacetate (2.0 mL, 20 mmol) in CH2C12 (25 mL) was treated with benzylamine (3.22 g, 30 mmol). The resulting mixture was stirred at room temperature for 16 h under an inert atmosphere of argon. Afterwards, the reaction was quenched with 2Ν HCl (10 mL). ' ,'The product was extracted with EtOAc (15 mL, 3X), dried over MgSO4, filtered and evaporated to an oil. The crude material was purified by flash chromatography using 9:1 hexane/EtOAc. , The product was isolated as a yellowish liquid (3.24 g, 86 % yield).
Η NMR (CDC13): δ 3.44 (s, 2H), 3.74 (s, 3H), 3.82 (s, 2H), 7.27 (m, IH), 7.34 (m, 4H).
Step B. Preparation of N-benzoyl-N-benzylgly cine
)
To a solution of N-benzylglycine methyl ester (800 mg, 4.5 mmol, step A) in THF (10 mL) was added benzoique acid (820 mg, 6.7 mmol) and DCC (1.40 g, 6.8 mmol). The reaction mixture was stirred at room temperature for 5 h. Then, it was treated with a saturated NaHCO3 solution and extracted with EtOAc (30 mL, 3X). The organic phase was evaporated to an oil.. The ester intermediate was saponified following the indications of example 65 (step E). The crude material was purified by flash chromatography using EtOAc:hexane:CHCl3 (5:5:2) to give 875 mg (73%) of the final product.
ΗNMR (DMSO-d6): δ 3.84 (s, 0.66H), 4.20 (s, 1.33H) ; 4,64 (s, 1.33H), 4.82 (s, 0.66H), 7.21 (d,7= 7.4, 2H), 7.40 (m, 8H), 7.55 (d, 7= 7.2, 2H), 10.0 (s, IH). LC-MS: 270 (M + H)+ and 292 (M + Na)+, 98% pure.
Step C. Preparation of (25) 2-N-isobu1 l-2-N-(4-methylberιzenesulfonyl)-6-N-(N'α-benzoyl-N'α- benzylglycyl)-2,6-diaminohexanol
. i • '
The title product was prepared from (25) 2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6- diaminohexanol (350 mg, 1.0 mmol, example 1.16, step A) following the indications of general procedure Ba using N-benzoyl-N-benzylglycine (269 mg, 1.0 mmol, step B) and NN- carbonyldiimidazole (180 mg, 1.1 mmol). The crude material was purified by preparative HPLC. The product was isolated as a solid (98 mg, 17% yield).
ΗΝMR (DMSO-d6): 5 0.83 (t,7= 7.1, 6H), 1.05 (m, 2H), 1.26 (m, 3H), 1.55 (m, IH), 1.87 (m, IH), 2.37 (s, 3H), 2.80-3.00 (m, 4H), 3.52 (m, IH), 3.65 (s, 0.66H), 3.89 (s, 0.33H), 4.49 (s, 0.33H), 4.66 (s, 0.66H), 7.20 (m, IH), 7.29 (m, 3H), 7.36 (d, 7= 7.3, 2H), 7.43 (m, 5H), 7.67 (d, 7= 7.5, 2H), 7.83 (m, IH). LC-MS: 594 (M + H)+, 99% pure.
Example 159. Preparation of (25) 2-N-isoburyi-2-N-(4-methylbenzenesulfonyl)-6-N-[N,α- benzyl-N'α-(4-methylbenzenesulfonyl)glycyl]-2,6-diaminohexanol
Step A. Preparation of N-(4-methylbenzenesulfonyl)-N-benzylglycine
A solution fo N-benzylglycine methyl ester (1.00 g, 5.6 mmol, example 158, step A) in DCM (10 mL) was treated with triethylamine (1 mL, 7.2 mmol) and 4-methylbenzenesulfonyl chloride (1.28 g, 6.7 mmol). The reaction mixture was stirred for 3 h at room temperature. Afterwards, the reaction was quenched with 2N HCl (10 mL). The product was extracted with DCM (15 mL, 3X), dried over MgSO4, filtered and evaporated. The ester intermediate was saponified following the indications of example 65 (step E). The crude material was diluted with 0.5N HCl (150 mL). The resulting precipitate was filtered off and dried under vacuum to give 1.46 g (82%) of the desired material (99%o pure). . .
ΗNMR (DMSO-d6): δ 2.40 (s, 3H), 3.80 (s, 2H), 4.40 (s, 2H), 7.21. (d, 7= 7.2, IH), 730 (m, 3H), 7.40 (d, 7= 7.6 2H), 774 (d, 7= 8.2, 2H),.12.7 (s, IH).
Step B . Preparation of (25) 2-N-isobutyl-2-N- (4-methylbenzenesulfonyl)-6-N-[N'α-benzyl-N'α- (4-methylbenzenesulfonyl)glycyl]-2,6-diaminohexanol
The title product was prepared from (25) 2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6- diaminohexanol (350 mg, 1.0 mmol, example 116, step A) following the indications of general procedure Ba using N-(4-methylbenzenesulfonyl)-N-benzylglycine (320 mg, 1.0 mmol, step A) and NN-carbonyldiimidazole (180 mg, 1.1 mmol). The crude material was purified by preparative HPLC. The product was isolated as a solid (236 mg, 37% yield).
Η ΝMR (DM'SO-d : δ 0.83 (t, 7= 7.3, 6H), 1.05 (m, 2H), 1.12 (m, 2H), 1.23 (m, IH), 1.55 (m, IH), 1.85 (m, IH), 2.36 (s, 3H), 2.40 (s, 3H), 2.80-3.00 (m, 4H), 3.34 (m, 2H), 3.52 (m IH), 3.65 (s, 2H), 439 (s, 2H), 7.25 (d, 7= 7.0, IH), 7.27-7.40 (m, 7H), 7.60 (t, 7= 4.2, IH), 7.66 (d, 7= 8.2, 2H), 7.75 (d, 7= 7.5, 2H). LC-MS: 644 (M + H)+, 90% pure.
Example 160. Preparation of (2S) 2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)- -N-(N'α- phenethyl-N'α-phenylglycyI]-2,6-diaminohexanol
Step A. Preparation of 2-phenylethyl-4-methylbenzenesulfonate
A solution of 2-phenylethanol (1.22 g, 10 mmol) in THF (10 mL) was added to a suspension of ΝaH (440 mg, 11.0 mmol, 60% in oil) in dry THF (5 mL) and stirred for 15 min. The resulting alcolate solution was added dropwise to a solution of 4-methylbenzenesulfonyl chloride (3.80 g, 20.0 mmol) in dry THF (10 mL) and stirred for a period of 1 h. Afterwards, the reaction was quenched with 2N HCl (5 mL). The product was extracted with EtOAc (25 mL, 3X), dried over MgSO4, filtered and evaporated to an oil. The crude material was purified by flash cliromatography using 9:1 hexane/EtOAc. The product was isolated as an oil (1.77 g, 64% yield).
Η MR (CDC13): δ 2.43 (s, 3H), 2.96 (t, 7= 73, 2H), 4.23 (t, 7= 7.2, 2H), 7.12 (d, 7= 7.3, 2H), 7.26 (m, 5H), 7.70 (d, 7= 7.6, 2H).
Step B. Preparation of N-phenethyl-N-phenylglycine
The title compound was prepared from N-phenylglycine (800 mg, 5.30 mmol) as described for the preparation of N-benzyl-N-phenylglycine .(example 114, step A) using 2-phenylethyl-4- methylbenzenesulfonate (1.61 g, 5.81 mmol) instead of benzylbromide. The crude material was purified by flash chromatography using a solvent gradient from 19:1 to 9:1 CH2Cl2/MeOH. The product was isolated as a solid (550 mg, 41% yield).
Η ΝMR (CDCI3): δ 2.95 (t, 7= 7.5, 2H), 3.65 (t, 7= 7.5, 2H), 3.95 (s, 2H), 6.73 (d, 7= 8.1, 2H), 6.80 (t, 7= 7.0, IH), 7.30 (m, 7H).
Step C. Preparation of (25) 2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-ό'-N-(N'α-phenethyl- ' N'α-phenylglycyl]-2,6.-diaminohexanol
The title product was prepared from (25) 2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6- diaminohexanol (320 mg, 0.94 mmol, example 116, step A) following the indications of general procedure Ba' using N-phenethyl-N-phenylglycine (200 mg, 1.0 mmol, step B) and NN- carbonyldiimidazole (140 mg, 0.86 mmol). The crude material was purified by preparative HPLC. The product was isolated as a hardy oil (32 mg, 7% yield).
ΗΝMR (DMSO-d6): δ 0.83 (t,7= 7.1, 6H), 1.05 (m, 2H), 1.25 (m, 3H), 1.55 (m, IH), 1.85 (m, IH), 2.37 (s, 3H), 2.80-3.01 (m, 4H), 3.52 (m, IH), 3.94 (s, 2H), 4.62 (s, 2H), 4.65 (t, 7= 4.3, IH), 6.58 (d, 7= 7.8, 2 H), 6.61 (t, 7= 7.8, 1 H), 7.11 (t, 7= 7.3, 2H), 7.25 (m, 3 H), 731 (t, 7 = 7.3, 2H), 735 (d, 7= 8.2, 2H), 7.66 (d, 7= 7.9, 2H), 7.81 (t, 7= 5.4, IH). LC-MS: 579 (M + H)+, 90% pure.
Example 161. Preparation of (2S,2'S) 2-N-isobutyl-2-N-(4-methylbenzenesulfonyI)-6-N- [7Vα-(4-aminobenzenesulfonyl)-S-tryptophanyl]-2,6-diaminohexanol
The title product was prepared from (25) 2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6- diaminohexanol (example 116, step A) following the indications of general procedure Bd using Nα~(4-nitroberιzenesulfonyl)-L-tryptophan (example 5, step A). The intermediate was reduced by catalytic hydrogenation following the conditions of general procedure E. Purification by HPLC gave 30 mg (14%) of the desired material.
]H ΝMR (CDC13): δ 0.84 (d, 7= 6.3, 3H), 0.89 (d, 7= 6.3, 3H), 0.94-1.03 (m, IH), 1.09-1.16 (m, 2H), 1.46-1.49 (m, IH), 1.93-1.98 (m, IH), 2.39 (s, 3H), 2.82-3.15 (m, 6H), 3.51 (d, 7= 6.8, 2H), 3.64 (q, 7= 63, IH), 3.89 (q, 7= 5.5, IH), 6.48 (d, 7= 7.8, 2H), 6.95 (t, 7= 4.5, IH), 7.19 (t, 7= 4.5, IH), 7.23-7.31 ( , 6H), 7.42 (t, 7= 4.5, IH), 7.73 (d, 7= 7.8, 2H).
Example 162. Preparation of (2S, S) 2-N-isobutyl-2-N-(4-methylbenzenesuIfonyl)-6-N- fN'α-(4-methoxyphenylaceryl)-S,-phenylalanyl]-2,6-diaminohexaιιol
The title product was prepared from (25) 2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6- diaminohexanol (example 116, step A) following the indications of general procedure Bd using Nα-(4-methoxyphenylacetyl)-L-phenylalanine (example 132, step A). Purification by HPLC gave 25 g (13%) of the desired material.
Η ΝMR (CDC13): δ 0.79 (d, 7= 6.3, 3H), 0.82 (d, 7= 6.3, 3H), 1.09-1.16 (m, 2H), 1.35-139 (m, 2H), 1.45-1.52 (m, IH) 1.94-1.97 ( , IH), 2.40 (s, 3H), 2.82-3.18 (m, 6H), 3.45 (s, 2H), 3.51 (d, 7= 7.0, 2H), 3,64 (q, 7= 6.3, IH), 3.79 (q, 7= 6.9, IH), 3.82 (s, 3H), 4.50 (q, 7= 5.6, IH), 6.87 (d, 7= 6.9, 2H), 6.89 (d, 7= 7.0, 2H), 7.15-7.21 (m, 3H), 7.35 (d, 7= 7.1, 2H), 7.50 (d, 7= Jl, lH), 7J3 (d, 7= 8.1, 2H). Example 163. Preparation of Nα-^-ammobenzenesulfony -Nα-isobutyl-Ne-ITV^α-^- methoxyphenylacety -L-phenylalanylj-L-lysine
The title compound was prepared from Nα-isobutyl-Nα-(4-nitrobenzenesulfonyl)-L-lysine hydrochloride (200 mg, 0.45 mfnol, example 5, step C) as described in general procedure Be using Nα-(4-methoxyphenylacetyl)-L-phenylalanine (example 132, step A). The final product, Nα-isobutyl-Nα-(4-rritrpbenzenesulfonyl)-Ne-[Nα-(4-methoxyphenylacetyl)-L-phenylalanyl]-L- lysine was subsequently hydrogenolysed following the indications of general procedure E. Purification by HPLC gave the desired material (7 mg, 4%).
Η ΝMR (CDC13): δ 0.79 (d, 7= 6.3, 3H), 0.82 (d, 7= 6.3, 3H), 1.09-1.16 (m, 2H), 1.35-1.39 (m, 2H), 1.45-1.52 (m, IH), 1.94-1.97 (m, IH), 2.40 (s, 3H), 2.82-3.18 (m, 6H),-3.45 (s, 2H), 3.64 (q, 7= 6.3, IH), 3.82 (s, 3H), 4.33 (t, 7= 5.5, IH), 4.50 (q, 7= 5.6, IH), 6.87 (d, 7= 6,9, 2H), 6.89 (d, 7= 7.0, 2H), 7.15-7.21 (m, 3H), 7.35 (d,7= 7.1, 2H), 7.50 (d,7= 7.1, IH), 7.73 (d, 7= 8.1, 2H).
Example 164., Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-[7Vα-(4- methoxyphenylacetyI)-L-tryptophanyl]-L-Iysine
Step A. Preparation of Nα-(4-methoxyphenylacetyl)-L-tryptophan '
L-tryptophan was reacted with 4-methoxyphenylacetyl chloride under the conditions used in general procedure A giving the title compound which was recrystallised from DCM (31 %) and used as such in the next step.
Step B. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-[N'α-(4- methoxyphenylacetyl)-L-tryptophanyl]-L-lysine
>
The title compound was prepared from Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine . hydrochloride (100 mg, 0.25 mmol, example 1, step E) -as described in general procedure Be using Nα-(4-methoxyphenylacetyl)-L-tryptophan (90 mg, 0.25 mmol) which was prepared in step A of this example. The final product was triturated with ether to yield 21 mg (11%) of the desired material.
I .
(
ΗNMR (CDC13): δ 0.79 (d, 7= 6.3, 3H), 0.82 (d, 7= 6.3, 3H), 1.09-1.16 (m, 2H), 135-139 ( , 2H), 1.94-1.97 (m, 2H), 2.30 (s, 3H), 2.82-3.18 (m, 6H), 3.45 (s, 2H), 3.82 (s, 3H), 4.25-4.20 (m, IH), 4.50 (q, 7= 5.6, IH), 6.77 (d, 7= 6.2, 2H), 6.89 (s, IH), 6.97 (d, 7= 6.9, 2H), 7.00-7.11 (m, 3H), 7.15-7.21 (m, 3H), 735 (d, 7= 7.1, 2H), 7.50 (d, 7= 7.1, IH), 7.73 (d, 7= 8.1, 2H).
Example 165. Preparation of (25,2'S) 2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N- [7Vα-(4-methoxyphenylacetyl)-S-tryptophanyl]-2,6-diaminohexanol
The title product was prepared from (25) 2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-256- diaminohexanol (example 116, step A) following the indications of general procedure Bd using Nα-(4-methoxy'phenylacetyl)-L-tryptophan (example 164, step A). Purification by HPLC gave 20 mg (10%) of the desired material.
Η ΝMR (CDC13): δ 0.79 (d, 7= 6.3, 3H), 0.82 (d, 7= 6.3, 3H), 1.09-1.16 (m, 2H), 135-139 (m, 2H), 1.94-1.97 (m, 2H), 2.30 (s, 3H), 2.82-3.18 (m, 6H), 3.45 (s, 2H), 3.51 (d, 7= 7.0, 2H), 3.82 (s, 3H), 4.50 (q, 7= 5.6, IH), 6.77 (d, 7= 6.2, 2H), 6.89 (s, IH), 6.97 (d, 7= 6.9, 2H), 7.00- 7.11 (m, 3H), 7.15-7.21 (m, 3H), 7.35 (d, 7= 7.1, 2H), 7.50 (d, 7= 7.1, IH), 7.73 (d,7= 8.1, 2H).
Example 166. Preparation of (2S,2'S) 2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-6-N- [7V α-(2-thiophenesulfonyl)-S-tryptophanyl]-2,6-diaminohexanol
Step A. Preparation of Nα-isobutyl-Nα-(4-acetamidobenzenesulfonyl)-L-α-amino-e-caprolactam
Nα-isobutyl-L-α-amino-e-caprolactam (4.14 g. 21.1 mmol, free base, example 1, step C) was dissolved in DCE (50 mL) and treated with diisopropylethylamine (6 mL, 0.3 mol) followed by
.) freshly recrystaUized 4-acetamidobenzenesulfonyl chloride (5.06 g, 21.6 mmol). The mixture was stirred overnight (TLC shows the reaction to be complete after 2 h). The solution was extracted with IN HCl (50 mL) and the organic layer was dried and evaporated. The crude material (7.01 g, 83%) was of sufficient purity to be used as such in the next step.
ΗNMR(CDC13): δ 0.93 (d,7=6.0, 3H), 0.96. (d,7= 6.0, 3H), 1.39 (t, 7= 12.0, IH), 1.85-1.65 (m, 3H), 2.08-2.18 (m and s, 6H), 2.90-2.97 (m IH), 3.00-3.06 (m, 2H), 3.35 (dd, 7= 14.2, 8.5, IH), 4.65 (d, 7= 8.7, IH), 6.3 (s, IH), 7.42 (d, 7= 8.8, 2H), 7.6 (d, 7= 8.8, 2H). '
Step B. Preparation of Nα-(4-aminobenzenesulfonyl)-Nα-isobutyl-L-lysine potassium salt
A mixture of Nα-isobutyl-Nα-(4-acetamidobenzenesulfonyl)-L-α-amino-e-caprolactam (6.8 g. 2 mmol) and, 6Ν HCl (200 mL) was refluxed for 12 h until all solids had disappeared. Afterwards, the solution was evaporated to dryness. The resulting solid was dissolved in EtOH (15 mL), neutralized with KOH and precipitated from acetone to give 8.0 g (100%) of the pure potassium salt. . ! ' ; ■
'HNMR (DMSO-d6): δ 072 (dd,7= 5.8, 6.4, 6H), 1.13-1.27 ( , 3H), 137-1.44 (ni IH), 1.72- lJ8 (m, IH), 1.92-1.98 (m, IH), 2.67-2.73 (m, 2H), 2.80-2.91 (m,2H), 3.85 (t,7=7.2, 1H), 6.56 (d, 7= 8.5, 2H), 7.44 (d, 7= 8.5, 2H).
Step C. Preparation of (25) 2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-2,6-diaminohexanol
A solution of Nα-(4-aminobenzenesulfonyl)-Nα-isobutyl-L-lysine (13.0 g, 40.0 mmol, step B) dissolved in MeOH (200 mL) was treated with of trimethylsilyl chloride (25 mL). The mixture was refluxed 3 h before stirring at room temperature for 2 h. Afterwards, the solution was evaporated and placed under high vacuum until a white solid was obtained (14.0 g). This was suspended in dry THF (100 mL) and added dropwise to a solution of LiAlH4 (5.0 g, 150 mmol) in THF (300 mL). The solution was stirred for 4 h. After cooling in an ice bath the solution was quenched by addition of MeOH (50 mL), water (5 mL), then 10%> ΝaOH (5 mL). The solvent
) was evaporated and the product was extracted from the precipitate with MeOH using a Soxlet apparatus during 18 h. Then, the solvent was evaporated to form a white solid which was dissolved in EtOH, filtered to eliminate Al2O3 and, after cooling, crystallized on standing (12.0 a 88%).
ΗNMR (DMSO-d6): δ 0.82 (m, 6H), 0.97-1.12 (m, 2H), 1.15-130 (im, 3H), 1.57 (m, IH), 1.84 (m, IH), 2.40 (t, 7= 7.0, 2H), 2.75 (m, IH), 2.85 (m, IH), 3.21 (m, IH), 3.44 (d, 7= 6.0, 2H), 5.92 (s, 2H), 6.59 (d, 7= 8.0, 2H), 7.39 (d, 7= 8.0, 2H).:
Step D. Preparation of (2S,2'S) 2-N-(4-aminobenzenesulfonyl)-2-N-isobutyI-6-N-[N'α-(2- thi oph enesulfonyl)-S-tryptophanyl] -2,6-diaminohexanol
The title product was prepared from (2S)- 2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-2,6- diaminohexanol (step C) following the indications of general procedure Bd using Nα-(2- thiophenesulfonyl)-L-tryptophan (example 53, step A). Purification by HPLC gave 200 mg (98%) of the desired material.
Η ΝMR (CDC13): δ 0.84 (d, 7= 6.3, 3H), 0.89 (d, 7= 6.3, 3H), 0.94-1.03 (m, IH), 1.09-1.16 (m, 2H), 1.46-1.49 (m, IH), 1.93-1.98 ( , IH), 2.82-3.15 (m, 6H), 3.51 (d,7= 6.8, 2H), 3.64 (q, 7= 6.3, IH), 4.05 (t, 7= 7.2, IH), 676 (d, 7= 6.5, 2H), 6.89 (t, 7= 4.6, IH), 6.97 (s, IH), 7.03 (t,7= 4.5, IH), 7.19 (t,7= 4.5, 1 H), 7.23-7.28 (m, 3H), 734-7.42 (m, 4H), 7.73 (d, 7= 6.8, 2H).
Example 167. Preparation of (2S,2'S) 2-N-(4-aminobenzenesuIfonyI)-2-N-isoburyI-6-N- (7Vα-benzenesulfonyl-S-try'ptophanyl)-2,6-diaminohexanol
The title product was prepared from (2S) 2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-2,6- diaminohexanol (example 166, step C) following the indications of general procedure Bd using Nα-benzenesulfonyl-L-tryptophan (example 4, step A). Purification by HPLC gave 196 mg (96%) of the desired material.
ΗΝMR (CDC13): δ 0.84 (d 7= 6.3, 3H)50.89 (d, 7= 6.3, 3H), 0.94-1.03 (m, IH), 1.09-1.16 (m, 2H), 1.46-1.49 ( , 1H),: 1.93-1.98 (m, IH), 2.82-3.15 (m, 6H), 3.51 (d, 7= 6.8, 2H), 3.64 (q, 7 = 6.3, IH), 3.94 (q, 7= 5.5, IH), 6.76 (d, 7= 6.5, 2H), 6.95 (t, 7= 4.5, IH), 7.19 (1, 7= 4.5, IH), 7.23-731 (m, 6H), 7.42 (t, 7= 4.5, IH), 7.60 (d, 7= 6.8, 2H), 7.73 (d, 7= 6.8, 2H).
Example 168. Preparation of (2S,2'S) 2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-6-N- (7Vα-aceryl-S-tryptophanyl)-2,6-diaminohexanol
The title product was prepared from (25) 2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-2,6- diaminohexanol (example 166, step C) following the indications of general procedure Bd using Nα-acetyl-L-tryptophan (example 99, step A). Purification by HPLC gave 165 mg (96%o) of the desired material.
!H ΝMR (CDC13): δ 0.90-0.98 (m, 6H), 1.09-1.16 ( , IH), 1.25-1.29 ( , IH), 1.45-1.52 (m, IH) 1.92-1.97 (m, IH),' 2.0, (s, 3H), 2.38 (s, 3H), 2.82-3.10 (m, 4H), 3.11-3.26 (m, 3H), 3.51 (d, 7= 7.0, 2H), 3i64 (q, 7= 6.3, IH), 3.75 (br s, IH), 6.86 (d, 7= 5.5, 2H), 6.77 (t, 7= 5.1, IH), 6.95 (t, 7= 5.2, 1H), 6.99 (s, 1 H), 7.15 (d, 7= 6.9, IH), 7.31 (d, 7= 7.1, IH), 7.45 (d, 7= 6.7, IH).
Example 169. Preparation of (2S,2'S) 2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-6-N- (7Vα-pivaloyl-S-tryptophanyl)-2,6-diaminohexanol
The title product was prepared from (25) 2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-2,6- diaminohexanol (example 166, step C) following the indications of general procedure Bd using Nα-pivaloyl-5-tryptophan (example 100, step A). Purification by HPLC gave 185 mg (97%o) of the desired material.
Η ΝMR (CDC13): δ 0.90-0.98 (m, 6H), 1.09-1.16 (m, IH), 1.23 (s, 9H), 1.25-1.29 (m, IH), 1.45-1.52 (m, IH), 1.92-1.97 (m, IH), 2.82-3.10 (m, 4H), 3.11-3.26 (m, IH), 3.41-3.45 (m, IH), ■ 4.51 (q, 7= 5.0, IH), 4.64 (br s, IH), 6.86 (d, 7= 5.5, 2H), 6.77 (t, 7= 5.1, IH), 6,85 (t, 7= 5.2, IH), 6.99 (s, IH), 7.15 (d, 7= 6.9, IH), 7.31 (d, 7= 7.1, IH), 7.45 (d, 7= 67, IH). Example 170. Preparation of (2S,2'S) 2-N-(4-aminobenzenesulfonyl)-2-N-isoburyl-6-7V- [7V α-(4-morpholinecarbohyl)-S-tryptophanyl]-2,6-diaminohexanol
Step A. Preparation of Nα-(4-morpholinecarbonyl)-L-tryptophan
L-tryptophan was reacted with 4-morpholinecarbonyl chloride under the conditions used in general procedure A giving the title compound as a thick oil. This material was used without further purification in the next step.
Step B. Preparation of (2S,2'S) 2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-6-N-[Nα-(4- morpholinecarbonyl)-S-tryptophanyl]-2,6-diaminohexanol
The title product was prepared from (25) 2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-2,6- diaminohexanol (example 166, step C) following the indications of general procedure Bd using Nα-(4-morpholinecarbonyl)-L-tryptophan (step A). Purification by HPLC gave 185 mg (97%) of the desired material. ■ '
Η ΝMR (CDC13): δ 0,90-0.98 ( , 6H), 1.09-1.16 (m, IH), 1.23 (5, 9H), 1.25-1.29 (m, IH), .1.45-1.52 (m, IH), 1.92-1.97 (m, IH), 2.82-3.10 (m, 4H), 3.11-336 (m, 6H), 3.41-3.55 (m, 4H), 4.14 (brs, IH), 6.86 (d,7= 5.5, 2H), 6.77 (t,7= 5.1, IH), 6.85 (t, 7= 5.2, IH), 6.99 (s, IH), 7.15 (d, 7= 6.9, IH), 7.31 (d, 7= 7.1, 'lH), 7.45 (d, 7= 6.7, IH).
Example 171. Preparation of (2S,2'S) 2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N- [7Vα-(4-morpholinecarbonyl)-S-tryptophanyl]-2,6-diaminohexanoϊ
The title product was prepared from (25) 2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6- diaminohexanol (example 116, step A) following the indications of general procedure Bd using Nα-(4-moφholinecarbonyl)-L-tryptophan (example 170, step A). Purification by HPLC gave 110 mg (57%) of the desired material. Η NMR (CDCI3): δ 0.90-0.98 (m, 6H), 1.09-1.16 ( , IH), 1.23 (s, 9H), 1,25-1.29 (m, IH), 1.45-1.52 (m, IH), 1.92-1.97 (m, 1H), 2.33 (s, 3H), 2.82-3.10 (m, 4H), 3.11-336 (m, 6H), 3.41- 3.55 (m, 4H), 4-14 (br s, IH), 6.86 (d, 7= 5.5, 2H), 6.77 (t, 7= 5.1, IH), 6.85 (t, 7= 5.2, IH), 6.99 (s, 1H), 7.15 (d, 7= 6.9, 1H), 731 (d, 7= 7.1, 1H), 7.45 (d, 7= 6J, IH).
Example 172. Preparation of (2S,2'S) 2-N-isobutyl-2-N-(4-methoxybenzenesuϊfonyI)-6-N- (7Vα-acetyl-S-tryptophanyI)-2,6-diaminohexanol
Treatment of (25,2'5)2-N-isobutyl-2-N-(4-methoxybenzenesulfonyl)-6-N-(S-tryptoρhanyl)-2,6- diaminohexanol trifluoroacetic acid salt (example 146) with acetyl chloride, using similar reaction conditions as for example 65 (step B) which uses 4-methylbenzenesulfonyl chloride, afforded the desired product in 86% yield.
1 , ' ! ' ' ' '• ' ' '
Η ΝMR (DMSO-d6): δ 0.82 (t, 7= 7.0, 6H), 1.02 (s, 9H), 1.00-1.30 (m, 5H), 1.77 (s, 3H), 2.80- 2.95 (m, 5H), 3.03 (dd, 7= 13.5, 7.0, IH), 3.77 (m, IH), 3.81 (s, 3H), 3.90 (m, IH), 3.98 (m, IH), 4.45 (m, IH), 6.96 (t, 7= 7.5, IH), 7.04 (t, j = 7.5, IH), 7.10 (d, 7= 8.0, 2H), 730 (d, 7= 7.5, IH), 7.58 (d, 7= 7.5, IH), 7.70 (d,7= 8.6, 2H),.7.85 (1, 7= 5.5, IH), 7.99 (d, 7= 8.5, IH), 1075 (s, IH).
Example 173. Preparation of (2S,2 'S) 2-N-cyclopentylmethyl-2-N-(4- methoxybenzenesuIfonyI)-6-N-(N'α-acetyl-S-tryptophanyl)-2,6- diaminohexanol
Treatment of (25,2'5) 2-N-cyclopentylmethyl-2-N-(4-methoxybenzenesulfonyl)-6-N-(5- tryptophanyl)-2,6-diaminohexanol trifluoroacetic acid salt (example 152) with acetyl chloride, using similar reaction conditions as for example 65 (step B) which uses 4-methylbenzenesulfonyl chloride, afforded the desired product in 76% yield.
'HΝMR (DMSO-d6): δ 0.93-130 (m, 7H), 1.40-1.68 (m, 7H), 1.77 (s, 3H), 2.18 (m, lH), 2.84- 3.10 (m, 5H), 3.30 (m, 2H), 3.50 (m, IH), 3.81 (s, 3H), 4.45 (m, IH), 4.65 (m, IH), 6.95-7.13 (m, 5H), 7.30 (d, 7= 8.3, IH), 7.56 (d, 7= 8.5, IH), 7.71 (d, 7= 8.2, 2H), 7.85 (t, 7= 5.5, IH), 7.98 (d, 7= 8.0, lH), 10.75 (s, lH). I '
Example 174. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-[7S^α-(4- methoxybenzyl)-7Vα-phenylglycyl]-L-lysine
Step A. Preparation of Nα-(4-methoxybenzyl)-Nα-phenylglycine
The title compound was prepared from N-phenylglycine (1.51 g, 10.0 mmol) as described for the preparation of N-benzyl-N-phenylglycine (example 114, step A) using 4-methoxybenzylchloride (1.72 g, 11.0 mmol) instead of benzylbromide. The crude material was purified by flash chromatography' using 9 :1 CH2Cl2/hexane. The product was isolated as ayellow solid (2.33 g, 86% yield). ' X
1H ΝMR (DMSO-d6): δ 3.72 (s, 3H), 4.11 (s, 2H), 4.51 (s, 2H), 6.59 (m, 3H), 6.87 (d, 7= 7.5, 2H), 7.11 (t, 7= 7.9! 2H), 7.21. (d, 7= 8.2, 2H), 12.6 (s, IH).
Step B. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-N6-[N'α-(4-methoxybenzyl)- Nα-phenylglycylJ-L-lysine
The title product was prepared from Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine hydrochloride' (360 mg, 1.0 mmol, example 1, step E) following the indications of general procedure Ba using Nα-(4-methoxybenzyl)-Nα-phenylglycine (271 mg, 1.0 mmol, step A) and NN-carbonyldiimidazole (195 mg, 1.2 mmol). The crude material was purified by preparative HPLC. The product was isolated as a solid (39 mg, 6%> yield).
Η ΝMR (DMSO-d6): δ 0.81 (t, 7= 6.9, 6H), 1.17 (m, 2H), 133. (m, 2H), 1.48 (m, IH), 1.80 (m, IH), 1.90 (m, IH), 237 (s, 3H), 2.85-3.05 (m, 4H), 3.71 (s, 3H), 3.90 (s, 2H), 4.16 (t, 7= 7.2, IH), 4.54 (s, 2H), 6.61 (t, 7= 8.7, 3H), 6.87 (d, 7= 8.4, 2H), 7.11 (t, 7= 7.4, 2H), 7.17 (d, 7= 8.6, 2H), 7.36 (d, 7= 7.6, 2H), 7.66 (d, 7= 8.3, 2H), 7.83 (t, 7= 5.1, IH), 12.75 (s, IH). LC- MS: 610 (M + H)+, 99% pure. Example 175. Preparation of (2S) 2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N-[7Vα- (4-methoxybenzyl)-7Vα-phenylglycyl]-2,6-diaminohexanol
The title product was prepared from (25) 2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6- diaminohexanol (345 mg, 1.0 mmol, example 116, step A) following the indications of general procedure Ba using Nα-(4-methoxybenzyl)-Nα-phenylglycine (271 mg, 1.0 mmol, example 174, step A) and NN-carbonyldiimidazole (195 mg, 1.2 mmol). The crude material was purified by preparative HPLC. The product was isolated as a yellow solid (105 mg, 18%) yield).
Η ΝMR (DMSO-d6): δ 0.82 (m, 6H), 1.01 (m, 2H), 1.23 (m, 3H), 1.48 (m, IH), 1.86 (m, IH), 2.36 (s, 3H), 2.85 (m, 1H), 2.95 (m, 3H), 3.26 (dd, 7= 11.2, 6.3, lHa), 332 (dd, 7= 11.2, 6.3, lHb), 3.52 (m, IH), 3.71 (s, 3H), 3.89 (s, 2H), 435 (m, IH), 4.54 (s, 2H), 6.61 ( , 3H), 6.86 (d, ' 7= 8.6, 2H), 7.11 (t, 7= 8.0, 2H), 7.17 (d, 7= 7.9, 2H), 7.36 (t, 7= 7.0, 2H), 7.66 (d, 7= 8.5, 2H), 7.78 (t, 7= 4.9, IH). LC-MS: 596 (M + H)+, 95% pure.
!
Example 176. Preparation of (25) 2-N-(4-aminobenzenesuIfonyl)-2-N-isobutyl-6-N-[7Vα- (4-methoxybenzyl)-7Vα-phenylglycyl]-2,6-diaminohexanol
The title product was prepared from (25) 2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-2,6- diaminohexanol (350 mg, 1.0 mmol example 166, step C) following the indications of general procedure Ba using Nα-(4-methoxybenzyl)-Nα-phenylglycine (271 nig, 1.0 mmol, example 17.4, step A) and NN-carbonyldiimidazole (195 mg, 1.2 mmol). The crude material was purified by preparative HPLC. The product was isolated as a yellow solid (87 mg, 14% yield).
]HΝMR (DMSO-d6): δ 0.81 (t,7= 7.0, 6H), 1.10 (m, 2H), 1.27 (m, 3H), 1.51 (m, IH), 1.85 (m, IH), 2.75 (dd,:7= 14.1, 7.3, lHa), 2.83 (dd,7= 14.1, 73, lHb), 2.98 (m, 2H), 3.22 (m, IH), 3.30 (m, IH), 3.45 (m, IH), 3.71 (s, 3H), 3.90 (s, 2H), 4.54 (s, 2H), 6.60.(m, 5H), 6,86 (d, 7= 8.8, 2H), 7.11 (1, 7= 7.6, 2H), 7.17 (d, 7= 7.7, 2H), 7.39 (d, 7= 8.7, 2H), 7.80 (t, 7 = 5.0, IH). LC- MS: 597 (M + H)+, 98% pure. Example 177. Preparation of (2S,2'S) 2-N-(4-aminobenzenesulfonyl)-2-7V-isobutyl-6-N- (7V,α-methoxtycarbonyl-S-phenylalanyl)-2,6-diamώohexanol
Step A. Preparation of Nα-methoxycarbonyl-L-phenylalanine | ,
L-phenylalanine was reacted with methyl chloroformate under the conditions used in general procedure A giving the title compound as an oil which was used without further purification in the next step. ' ι , ■ ' ,
Step B. Preparation of ; (25,2'5) 2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-6-N-(N,α- methoxycarbonyl-S-phenylalanyl)-2,6-diaminohexanol
The title product was prepared from (25) 2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-2,6- diaminohexanol (example 166, step C) following the indications of general procedure Bd using Nα-methoxycarbonyl-L-phenylalanine (step A). Purification by HPLC gave 159 mg (98%) of the desired material.
Η ΝMR (CDC13): δ 0.79 (d, 7= 6.3, 3H), 0.82 (d, 7= 63, 3H), 1.08-1.11 (m, 2H), 1.23-131 (m, 2H), 1.45-1.52 (m, IH), 1,92-1.97 (m, IH), 1.98 (d, 7= 9.0, 3H), 2.84-3.09 (m, 6H), 3.51 (d,7= 7.1, 2H), 3.59 (s, 3H), 3.61-3.64 (m, IH), 4.51 (q,7= 6.9, IH), 6.76 (d,7= 8.1, 2H), 7.09- 7.26 (m, 5H), 7.73 (d, 7= 8.1, 2H). LC-MS: '549.7 (M + H)+, 98% pure.
Example 178. Preparation of (2S,2'S) 2-N-(4-aminobenzenesuIfonyI)-2-N-isobutyl-6-N- (7Vα-methoxycarbonyl-S-try'ptophanyl)-2,6-diaminohexanol
Step A. Preparation of Nα-methoxycarbonyl-L-tryptophan
L-tryptophan was reacted with methyl chloroformate under the conditions used in general procedure A giving the title compound as an oil which was used without further purification in the next step. Step B. Preparation of (2S,2'S) 2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-6-N-(N'α- methoxycarbonyl-S-tryptophanyl)-2,6-diaminohexanol
The title product was prepared from (25) 2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-2,6- diamiriohexanol (example 166, step C) following the indications of general procedure Bd using Nα-methoxycarbonyl-L-tryptophan (step A). Purification by HPLC gave 155 mg (87%>) of the desired material.
'H ΝMR (CDC13): δ 0.90-0.98 (m, 6H), 1.09-1.16 (m, IH), 1.25-1.29 (m, IH), 1.45-1.52 (m, IH), 1.92-1.97 (m, IH), 2.82-3.10 (m, 4H), 3.11-3.26 (m, 3H), 3.51 (d,7=7.0, 2H), 3.59 (s, 3H), 3.64 (q, 7= 6.3, IH), 3.75 (br s, IH), 6.86 (d, 7= 5.5, 2H), 6.77 (t, 7= 5.1, IH), 6.95 (t,7= 5.2, lH), 7.15 (d,7= 6.9, 1H), 731 (d, 7= 7.1, IH), 7.45 (d, 7= 6.7, IH). LC-MS: 588.7 (M + H)+, 98% pure. .
Example 179. Preparation of (2S, S 2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-6-N- [7Vα-(4-nitrobenzenesulfonyl)-S-tryptophanyI]-2,6-diaminohexanol
The title product was prepared from (25) 2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl -2,6- diaminohexanol (example 166, step C) following the indications of general procedure Bd using Nα-(4-nitrobenzenesulfonyl)-L-tryptophan (example 5, step A). Purification by HPLC gave 255 mg (60%) of the desired material.
LC-MS: 715.8 (M + H)+, 98% pure. • ''
Example 180. . ' Preparation of (2S,2'S) 2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-6-N- [7Vα-(4-morpholinecarbonyl)-S;-phenylalanyI]-2,6-diammohexanol
The title product was prepared from (25) 2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-2,6- diaminohexanol (example 166, step C) following the indications of general procedure Bd using Nα-(4-morpholinecarbonyl)-L-phenylalanine (example 51, step A). Purification by HPLC gave 165 mg (91%) of the final product. Η NMR (CDC13): δ 0.79 (d, 7- 6.3, 3H), 0.82 (d, 7= 6.3, 3H), 1.08-1.11 (m, 2H), 1.23-131 (m, 2H), 1.45-1.52 ( , IH), 1.92-1.97 (m, IH), 2.84-3.09 (m, 6H), 3.21-331 (m, 4H),3.45-3.64 (m, 7H), 4.48 (br s, IH), 6.88 (d, 7= 8.1, 2H ), 7.09-7.26 (m, 5H), 7.43 (d, 7= 8.1, 2H). LC-MS: 604.8 (M + H)+, 98% pure. . . .
Example 181. Preparation of (25) 2-N-(4-aminobenzenesuIfonyI)^2-N-isobutyl-6-N-(7Vα- •i isobutyl-7Vα-phenylglycyl)-2,6-diaminohexanol
Step A. Preparation of N-isobutyl-N-phenylglycine i The title compound was prepared from N-phenylglycine (1.0 g, 5.5 mmol) as described for the preparation of N-benzyl-N-phenylglycine (example 114, step A) using isobutyl iodide (2.0 mL,
17.3 mmol) instead of benzylbromide. The crude material was purified by flash chromatography using hexane/EtOAc/CHCl3 (7:3:2) as the eluent. The product was isolated as a brown oil (560 mg, 41% yield).
'H ΝMR (CDC13): δ 0.94 (rn, 6H), 2.03 (m, IH), 3.16 (d, 7= 7.2, 2H), 4.06 (s, 2H), 6.66 (d, 7 = 8.2, 2H), 6.75 (t, 7= 7.2, IH), 7.21 (1, 7= 8.0, 2H).
Step B. Preparation of (25) 2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-6-N-(N'α-isobutyl-N'α- phenylglycyl)-2,6-diaminohexanol
A solution of N-isobutyl-N-phenylglycine (200 mg, 1.0 mmol, step A) in DMF (5.0 mL) was treated with EDC (290 mg, 1.5 mmol) and HOBt (100 mg, 0.75 mmol) for a period of 10 min under an inert atmosphere or argon. Then, (25) 2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-2,6- diaminohexanol (345 mg,T.O mmol example 166, step C) was added and the reaction mixture was stirred for a period of 6 h. Afterwards, a 1 % aqueous citric acid solution was added and the product extracted with EtOAc (10 mL, 3X). The organic phase was washed with water 10 mL (3X) and with brine (5 L). The organic phase was dried with MgSO4, filtered and evaporated to a solid. The crude material was purified by preparative HPLC. The product was isolated as a yellow solid (125 mg, 24% yield). 1 'HNMR (DMSO-d6): δ 0.81 (t, 7 = 6.5, 6H), 0.88 (d,7= 8.1, 6H), 1.05 (m, IH), 1.13 (m, IH), 1.26 (m, 3H), 1.55 (m, IH), 1.84 (m, IH), 1.98 (m, IH), 2.74 (dd, 7 = 13.6, 7.1, IH), 2.83 (dd, 7= 14.2, 7.5, IH), 2.95 (d, 7= 6.4, 2H), 3.18 (d, 7= 7.3, 2H), 3.23 (m, IH), 3,44 (m, IH), 3.85 (s, 2H), 4.59 (t, 7 = 5.0, IH), 5.90 (s, 2H), 6.58 (m, 5H), 7:11 (t, 7 = 7.4, 2H), 738 (d, 7 = 8.6, 2H), 7.67 (t, 7= 5.0," IH). LC-MS: 553 (M + H)+, 98% pure.
Example 182. Preparation of (25) 2-N-(4-aminobenzenesulfonyI)-2-N-isobutyl-6-N-(7Vα- benzyI-7Vα-phenyIglycyI)-2,6-diaminohexanoI
The title product was* prepared from (25) 2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-2,6- diaminohexanol (345 mg, 1.0 mmol, example 166, step C) as described above for the preparation of 2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-6-N-(N α-isobutyl-N α-phenylgly cyI)-2,6- diaminohexanol (example 183) using N-benzyl-N-phenylglycine (241 mg, 1.0 mmol, example 114, step A) instead of N-isobutyl-N-phenylglycine. , The crude material was purified by preparative HPLC. The product was isolated as a yellow solid (370 mg, 65%> yield).
!H ΝMR (DMSO-d6): δ^ 0.81 (t, 7= 6.9, 6H),.1.05 (m, IH), 1.13 (m, IH), 1.28 (m, 3H), 1.55 (m, IH), 1.84 (m, lH), 2J4 (dd, 7= 12.6, 7.0, 1H), 2.85 (dd, 7= 113, 5.9, 1H), 2.98 (d,7= 6.0, 2H), 3.24 (dd, 7= 11.7, 6.3, IH), 3.30 (dd, 7= 8.9, 5.0, IH), 3.45 ( , IH), 3.94 (s, 2H), 4.62 (s, 2H), 5.35 (br s 2H), 6.60 (m, 5H), 7.11 (t, 7= 7.7, 2H), 7.24 (m, 3H), 7.31 (t, 7= 7.5, 2H), 7.39 (d, 7= 7.6, 2H), 7.83 (t, 7= 5.1, IH). LC-MS: 567 (M + H)+, 99% pure.
Example 183. Preparation of (2S, S) 2-N-isobutyl-2-N-(4-methyIbenzenesulfonyl)-6-N-(S- tryptophanyl)-2,6-diaminohexanol trifluoroacetic acid salt
Step A. Preparation ofNα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-(Nα-tert-butoxycarbonyl- L-tryptophanyl)-L-lysine methyl ester
To a stirred solution of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-L-lysine methyl ester (370 mg, 1 mmol, example 65, step C) in THF/K2CO3 (IM) (3 mL/3 mL) was added Na-tert- butoxycarbonyl-L-tryptophan N-hydroxysuccinimide ester (550 mg, 1.2 mmol). The reaction mixture was stirred overnight then diluted with IN HCl and extracted with EtOAc. The organic layer was dried (MgSO4) and concentrated. The crude was purified by flash chromatography using hexane/EtOAc as eluent to afford the. desired product (85% yield). .
•
ΗΝMR(DMSO-d6): δ 0.81 (t,7=7.0, 6H), 1.10-1.50 (m, 5H), 1.31 (s, 9H), 1.75-1.90 (m,2H), 2.38 (s, 3H), 2.82-3.10 (m, 5H), 3.44 (s, 3H), 4.12 (m, IH), 4.30 (t,7= 5.0, IH), 6.67 (d, 7= 8.2, IH), 6.90-7.12 (m, 5H), 7.30 (d,7= 8.0, IH), 7.40 (d,7= 7.5, 2H), 7.55 (m, IH), 7.65.(d, 7= 7.5, 2H), 7.78 (m, IH), 10.77 (s, IH).
Step B. Preparation of (2S2'5) 2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N-(N'α-tert- butoxycarbonyl- 5,-tryptpphanyl)-2,6-diaminohexanol !
Nα-isobuty'l-Λ?'α-(4-methylbenzenesulfonyl)-N6-(Nα-tert-butoxycarbonyl-L-tryptophanyl)-L- lysine methyl , ester (step A) was reduced with LiAlH4 following the indications of general procedure G. The final product was obtained with 78% yield.
!HΝMR(DMSO-d5): δ 0.84 (t,7= 7.0, 6H), 0.90-1.27 (m, 5H), 1.31 (s, 9H), 1.52 (m, IH), 1.85 (m, IH), 2.37 (s, 3H), 2.80 and 3.02 (ABX, 7= 14.0, 7.2, 2H), 2.90 (rn, 2H), 3.33 (m, IH), 3.50 (m, IH), 4.15 (m, IH), 4.65 (t, 7= 5.0, IH), 6.65 (d, 7= 7.8, IH), 6.92-7.12 (m, 5H), 7.30 (d, 7 = 7.8, IH), 7.35 (d, 7= 7.5, 2H), 7.57 (d, 7= 8.0, IH), 7.67 (d, 7= 7.7, 2H), 7J3 (t, 7= 5.0,.1H), ιoJ7.(s, rn).': • , ; . . ' ■ ■ . i
!
Step C. Preparation ' of (2S,2'S) 2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N-(S- tryptophanyl)-256-diaminohexanol trifluoroacetic acid salt
This product was obtained quantitatively by treating (2S,2'S) 2-N-isobutyl-2-N-(4- memylbenzenesulfonyl)-6-N-(Nα-tert-butoxycarbonyl-S-tryptophanyl)-2,6-diaminohexanol (step B) with TFA in CH2C12. ΗN R CDMSO-dβ): δ 0.80 (d, 7= 7.0, 3H), 0.83 (d, 7= 7.0, 3H), 0.92-1.60 (m, 6H), 1.88 (m, IH), 2.37 (s, 3H), 2.78-3.00 (m, 4H), 3.05 and 3.20 (ABX, 7= 14.0, 7.0, 2H), 3.22 and 330 (ABX,7= 14.2, 7.0, 2H), 3.50 (m, IH), 3.86-3.97 (m, 2H),4.35 (m, IH), 6.95-7.12 (m, 4H), 7.18 (s, IH), 7.38 (d, 7= 8.2, IH), 7.60 (m, IH), 7.70 (d, 7= 8.2, 2H), 8.05 (br s, 3H), 8.34 (m, IH), 10.99 (s, IH).
Example 184. Preparation of (2S,2'S) 2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-6-N- (/Vα-isobutyryl-S-tryptophanyl)-2,6-diaminohexanol
Step A. Preparation of Nα-isobutyryl -L-tryptophan i ' ! ' ' ' :
L-tryptophan was reacted with isobutyryl chloride under the conditions used in general procedure A giving the title compound which was recrystallised from DCM (55%). This material was used without further purification in the next step.
Step B. Preparation of (2S,2'S) 2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-6-N-(Nα-isobutyryl- 5-tryptophanyl)-2,6-diaminohexanol
The title product was prepared from (25) 2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-2,6- diaminohexanol (example 166, step C) following the indications of general procedure Bd using Nα-isobutyryl-L-tryptophan (step A). Purification by HPLC gave 121 mg (67%) of the desired material. ' • . :
1HΝMR (DMSO-d6): δ 0.80-0.90 (m, 9H), 0.95-0.97 (d, 7= 6.6, 3H), 1.09-1.16 (m, IH), 1.25-
1.29 (m, IH), 1.45-1.52 (m, IH), 1.92-1.97 (m, IH), 233-2.45 (m, IH), 2.82-2.99 (m, 4H), 3.26- i
3.33 (m, 1H), 3.51-3.55 (m, IH), 4.64 (br s, lH), 6.66 (d,7= 5.5, 2H), 6.87 (t, 7= 5.1, 1H), 6.95 (t, 7= 5.2, IH), 6.99 (s,'l H), 7.15 (d, 7= 6.9, 1H), 731 (d, 7= 7.1, 1H), 7.45 (d, 7= 6.7, IH). LC-MS: 600.8 (M + H)+, 99% pure. Example 185. • Preparation of (2S,2'S) 2-7V-(4-aminobenzenesulfonyl)-2-N-isobutyl-6-N- (7Vα-tert-butylacetyl-S-tryptophanyl)-2,6-diaminohexanol
Step A. Preparation of Nα-te7't-butylacetyl-L-tryptophan
L-tryptophan was reacted with tert-butylacetyl chloride under the' conditions used in general procedure A giving the title compound which was recrystallised from DCM (81 %). This material was used as such in the next step.
Step B. Preparation of (2S,2'5) 2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-6-N-(Nα-tert- butylacetyl-S-tryptophanyl)-2,6-diaminohexanol . ;
The title product was prepared from (25) 2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-2,6- diaminohexanol (example 166, step C) following the indications of general procedure Bd using Nα-rert-butylacetyl-L-tryptophan (step A). Pμrification by HPLC gave 133 mg (70%) of the desired material.
!H ΝMR (DMSO-d6): δ' 0.80-0.90 (m, 15H), 1.09-1.16 (m, IH), 1.25-1.29 (m, IH), 1.45-1.52 (m, IH), 1.82-1.87 (m, IH), 1.97 (s, 2H), 233-2.45 (m, IH), 2.82-2.99 (m, 4H), 3.26-333 (m, IH), 3.51-3.55, (m, IH), 4.64 (br s, IH), 6.66 (d, 7= 5.5, 2H), 6.87 (t, 7= 5.1, IH), 6.95 (t, 7= 5.2, 1H),'6.99 (s, IH), 7.15 (d,7= 6.9, IH), 7.31 (d,7= 7.1, IH), 7.45 (d,7= 6J, IH). LC-MS: • 628.8 (M + H)+, 98% pure. <
Example 186. Preparation of (2S,TS) 2-N-(4-aminobenzenesuIfonyl)-2-N-isobutyI-6-N- (Λ^α-benzoyl-S-tryptophanyl)-2,6-diaminohexanol
Step A. Preparation of Nα-benzoyl-L-tryptophan
)
L-tryptophan was reacted with benzoyl chloride under the conditions used in general procedure
A giving the title compound which was recrystallised from DCM (51%). This compound was used without further purification in the next step. Step B . Preparation of (25,2'5) 2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-6-N-(N α-benzoyl- S-tryptophanyl)-2,6-diaminohexanol
The title product was prepared from (25) 2-N-(4-aminobenzenesμlfonyl)-2-N-isobutyl-2,6- diaminohexanol (example 166, step C) following the indications of general procedure Bd using ( Nα-benzoyl-L-tryptophan (step A). Purification by HPLC gave 122 mg (64%) of the desired material.
ΗΝMR (CDC13): δ 0.84(d,7= 63, 3FI), 0.89 (d,7= 63, 3H), 0.94-1.03 (m, IH), 1.09-1.16 (m, 2H), 1.46-1.49 (m, IH), 1.93-1.98 (m, IH), 2.39 (s, 3H), 2.82-3.15 (m, 6H), 3.51 (d,7=6.8, 2H), 3.64 (q, 7= 6.3, lH), 3.94 (q, 7= 5.5, lH), 6.95.(t, 7= 4.5, IH), 7.19 (t, 7= 4.5, IH), 7.23-731 ( , 6H), 7.42 (t, 7= 4.5, IH), 7.60 (d, 7= 6.8, 2H), 7.73 (d, 7= 6.8, 2H). LC-MS: 634.8 (M + H)+, 98% pure. : t
Example 187. Preparation of (2S, S) 2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-6-N- [7Vα-(4-aminobenzenesulfonyl)-S-tryptophanyl]-2,6-diaminohexanol
The title compound was obtained from (25,2'5) 2τN-(4-aminobenzenesulfonyl)-2-N-isobutyl-6-N- [Nα-(4-nitrobenzenesulfonyl)-5-tryptophanyl]-2,6-diaminohexanol (200 mg, 0.28 mmol, example 179) by catalytic, hydrogenation following the conditions of general procedure E. Purification by HPLC gave 191 mg (99%) of the desired material.
]HΝMR (DMSO-d6): δ 0.73 (d,7= 63, 3H), 0.75 (d,7= 63, 3H), 1.00-1.11 (m, 4H), 132-135 (m, IH), 1.66-1.69 (m, IH), 1.83-1.88 (m, 1H), 2.48 (br s, 6H), 2.59-2.67 (m, 2H), 2.84-2.96 (m, 2H), 3.20 (d, 7= 6.5, 2H), 4.21 (t, 7= 7.2, IH), 6.66 (d, 7= 7.1, 2H), 6.72 (d, 7= 7.1, 2H), 6.85 (t, 7= 4.0, IH), 7.09 (t, 7= 4.0, 2H), 7.28 (d, 7= 7.1 ,1H), 7.33 (d, 7= 7.1, 2H), 7.60 (t, 7= 4.0, IH). LC-MS: 685.8 (M +;H)+, 98% pure. Example 188. Preparation of (2S,2'S) 2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N- (7Vα-benzenesulfonyl-S-cyanoaIanyl)-2,6-diaminohexanol
The title product was prepared from (25) 2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6- diaminohexanol (example 116, step A) following the indications of general procedure Bd using Nα-benzenesulfonyl-L-asparagine (example 25, step A). In tins particular reaction an excess EDC (2.5 eq.) was used which dehydrated the amide function on the asparagine moiety. Purification by HPLC gave 6 mg (9%) of the desired material.
LC-MS: 579.2 (M + H)+, 92% pure. ; j
Example 189.. Preparation of Nα-isobutyl-Nα-(4-methyIbenzenesulfonyl)-Ne-(N,α- benzenesulfonyl-L-tryptophanyl)-L-lysine amide
A solution of Nα-isobutyl-Λ^α-(4-methylbenzenesulfonyl)-Nε-(N'α-benzenesulfonyl-L- tryptophanyl)-L-lysine (100 mg, 0.15 mmol, example 4) dissolved in EtOAc (3 mL) was treated with DCC (30 mg, 0.15 mmol) and N-hydroxysuccinimide (17 mg, 0.15 mmol). The resulting solution was stirred overnight at room temperature. Then, the reaction mixture was filtered through celite. The organic solvent was evaporated and the crude residue dissolved in THF (5 mL) was treated with concentrated ΝH4OH (1 mL). The reaction mixture was left in the refrigerator overnight. Afterwards, it was filtered through celite and the solvent was evaporated to give 98 mg (97%) of the desired title compound after purification by HPLC. .
LC-MS: 682.8 (M + H)\ 99% pure.
Example 190. ^ Preparation of (2S,TS) 2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N- (N^x-methoxycarbonyl-S-tryptophanyl)-2,6-diaminohexanol
The title product was prepared from (25) 2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6- diaminohexanol (example 116, step A) following the indications of general procedure Bd using Nα-methoxycarbonyl-L-tryptophan (example 178, step A). Purification by HPLC gave 21 mg (15%) of the desired material.
Η NMR (CDCI3): δ 0.90-0.98 (m, 6H), 1.09-1.16 (m, IH), 1.25-1.29 (m, IH), 1.45-1.52 (m, IH), 1.92-1.97 ( , IH), 238 (s, 3H), 2.82-3.10 (m,4H), 3.11-3.26 (m, 3H), 3.51 (d,7= 7.0, 2H), 3.59 (s, 3H), 3.64 (q,7= 6.3, IH), 3.75 (br s, IH), 6.77 (t,7= 5.1, IH), 6.95 (t,7= 5.2, IH), 6.99 (s, IH), 7.15-7.25 (m, 3H), 7.31 (d, 7= 7.1, IH), 7J5 (d, 7= 6.7, IH). LC-MS: 587 (M + H)+, 99% pure.
Example 191. Preparation of (2S, S) 2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N- (N'α-benzoyl-S-tryptophanyI)-2,6-diaminohexanol
The title product was prepared from (25) 2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6- diaminohexanol (example 116, step A) following the indications of general procedure Bd using Nα-benzoyl-L-tryptophan (example 186, step A). Purification by HPLC gave 41 mg (28%) of the desired material.
Η ΝMR (CDC13): δ 0.90-0.98 (m, 6H), 1.09-1.16 (m, IH), 1.25-1.29 (m, IH), 1.45-1.52 (m, IH), 1.92-1.97 (m, lH), 2.82-3.10 (m, 4H), 3.11-3.26 (m, 3H), 3.51 (d,7= 7.0, 2H), 3.59 (s, 3H), 3.64 (q.7= 6.3, IH), 4.08 (br s, IH), 6.86 (d, 7= 5.5, 2H), 6.77 (t, 7= 5.1, IH), 6.95 (t, 7= 5.2, IH). 6.99 (s, IH), 7.15 (d, 7= 6.9, IH), 731 (d, 7= 7.1, IH), 7.45 (d, 7= 6.7, IH). LC-MS:
633.8 (M + H)+, 99% pure.
1
Example 192. Preparation of (2S,2'5) 2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N- (N'α-isobutoxycarbonyl-S-tryptophanyl)-2,6-diaminohexanol
Step A. Preparation of Nα-isobutoxycarbonyl-L-tryptophan
L-tryptophan was reacted with isobutyl chloroformate under the conditions used in general procedure A giving the title compound which was recrystallised neat. This product was used without further purification in the next step. Step B. Preparation of (2S,2'S) 2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-6-N-(N'α- isobutoxycarbonyl-5-tryptophanyl)-2,6-diaminohexanol
The title product was prepared from (25) 2-N-isobutyl-2-N-(4-methylbenzenesulfonyl)-2,6- diaminohexanol (example 116, step A) following the indications of general procedure Bd using Nα-isobutoxycarbonyl-L-tryptophan (step A). Purification by HPLC gave 64 mg (40%) of the desired material.
,HΝMR (DMSO-d6): δ 0.80-0.91 (m, 9H), 0.94-0.97 (d,7= 6.6, 3H), 1.09-1.16 (m, IH), 1.25- , 1.29 (m, IH), 1.45-1.52 (m, IH), 1.92-1.97 ( , IH), 233-2.45 (m and s, 4H), 2.82-2.99 (m, 6H), 3.26-333 (m, IH), 3.51-3.55 (m, IH), 4.44 (br s, IH), 6.87 (t,7= 5.1, IH), 6.95 (t, 7= 5.2, IH), 6.99 (s, IH), 7.15-7.22 (m, 3H), 7.31 (d, 7= 7.1, IH), 7.65 (d, 7= 6.7, IH). LC-MS: 629.8 (M + H)+, 99% pure.
Example 193. Preparation of (2S, S) 2-Λ'-(4-ammobenzenesulfonyl)-2-N-isobutyl-6-N- (N'α-isobutoxycarbonyl-S-tryptophanyl)-2,6-diaminohexanol
The title product was prepared from (25) 2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-2,6- diaminohexanol (example 166, step C) following the indications of general procedure Bd using Nα-isobutoxycarbonyl -L-tryptophan (example 192, step A). Purification by HPLC gave 151 mg (95%) of the final product.
]HΝMR (DMSO-d6): δ10.80-0.90 (m, 9H), 0.95-0.97 (d, 7= 6.6, 3H), 1.09-1.16 (m, IH), 1.25- 1.29 (m, IH), 1.45-1.52 (m, IH), 1.92-1.97 (m, IH), 233!-2.45 (m, IH), 2.82-2.99 (m, 6H), 3.2.6- 3.33 (m, IH), 3.51-3.55 (m, IH), 4.44 (br s, IH), 6.66 (d, 7= 5.5, 2H), 6.87 (t, 7= 5.1, IH), 6.95 (t, 7= 5.2, IH), 6.99 (s, 1 H), 715 (d, 7= 6.9, IH), 7.31 (d, 7= 7.1, IH), 7.45 (d, 7=.6J, IH). LC-MS: 630.2 (M + H)+, 99% pure. Example 194. Preparation of (2S,2'S) 2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-6-N- [N,α-(4-nitrobenzenesulfonyl)-S-phenylalanyl]-2,6-diaminohexanol
The title product was prepared from (25) 2-N-(4-ammobenzenesulfonyl)-2-N-isobutyl-2,6- ■ diaminohexanol (example 166, step C) following the indications of general procedure Bd using Nα-(4-nitrobenzenesulfonyl)-L-phenylalanine (example 10, step A). Purification by HPLC of about half (50 mg) of the crude material gave 20 mg (40%) of the final product. i . I '
LC-MS: 676.8 (M + H)+, 95% pure.
P ' < x ■ x ' i . • :
Example 195. ; Preparation of (2S,2'S) 2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-6-N- [N,α-(4-aminobenzenesulfonyl)-S-phenylalanyl] -2,6-diaminohexanol
The title compound was obtained from (25,2'5) 2-N-(4-aminobenzenesulfonyl)-2-N-isobutyl-6-N- [N'α-(4-nitrobenzenesulfonyl)-S-phenylalanyl]-2,6-diaminohexanol (50 mg, 0.07 mmol, example 194) by catalytic hydrogenation following the conditions of general procedure E. Purification by HPLC gave 31 mg (77%) of the desired material.
LC-MS: 646.8 (M + H)+, 99% pure. j ' • X
Example 196.; Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyI)-Ne-[N'α-(4- I methoxyanilinecarbonyl)-L-phenylalanyl]-L-lysine
The title compound ! was prepared from solid phase bound Nα-isobutyl-Nα-(4- methylbenzenesulfonyl)-Ne-(9-fluorenylmethoxycarbonyl)-L-lysine (450 mg, 0.125 mmol) as described in general . procedure Bb • using commercially available Nα-(9- fluorenylmethoxycarbonyl)-L-phenylalanine (400 mg, 0.9 mmol). After the coupling reaction, the resin was once more deprotected and activated with NN-carbonyldiimidazqle (5-10 fold excess) for 30 min. after which time the resin was washed with DCM (4X) before 4- methoxy aniline was added. The tube was sealed and left for a period of 12 h. Afterwards, the product was cleaved from the resin using TFA as indicated in general procedure Bb. The final product was purified by preparative HPLC to yield 11 mg (13%o) of the desired material.
ΗNMR (CDC13): δ 0.83 (d,7= 6.9, 6H), 1.08-1.11 (m, 2H), 133-1.55 (m, 2H), 1.45-1.52 (m,, IH), 1.79-1.89 (m,,2H), 2.36 (s, 3H), 2.85-3.27 (m, 6H), 3.55 (s, 2H), 3J9 (s, 3H), 4.21 (s, 2H), 433 (t,7= 4.5, IH), 6.69 (d,7= 8.2, 2H), 6.99-7.19 (m, 3H), 7.15-7.26 (m, 7H), 7J3 (d,7= 8.1, 2H). LC-MS: 651.8 (M - H)\ 99% pure.
Example 197.' Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-Ne-(N,α- i pyrrolidinecarbonyl-L-phenylalanyl)-L-lysine
. • i This compound was prepared as described for the preparation of Nα-isobutyl-Nα-(4- memylbenzenesulfonyl)-N6-[N'α-(4-methoxyanilinecarbonyl)-L-phenylalanyl]-L-lysine (example
196) using pyrrolidine instead of 4-methoxyaniline. The crude .material was purified by preparative HPLC to give 15 mg, 20% of the desired material. .
LC-MS: 599.7 (M - H)", 99%' pure.
Example 198.; Preparation of Nα-isobutyl-Nα-(4-methylbenzenesuIfonyI)-Ne-(N'α- methyIaminocarbonyl-L-phenylalanyl)-L-lysine
This compound was prepared as described for the preparation of Nα-isobutyl-Nα-(4- methylbenzenesulfonyl)-Ne-[N'α-(4-memoxyamlinecarbonyl)-L-phenylalanyl]-L-lysine (example 196) using methylamine instead of 4-methoxyaniline. The crude material was purified by preparative HPLC to give 4 mg, 6% of the desired material.
ΗΝMR O DCl,): δ 0.83 (d, 7= 6.9, 6H), L08-l.l l (m, 2H), 1.23-1.25 (m, 2H), 1.45-1.52 (m, IH), 1.89-1.99 (m, 2H), 2.32 (s, 3H), 2.70 (s, 3H), 2.94-3.09 (m, 6H), 4.23 (t, 7= 5.9, IH), 4.61 (m, IH), 7.09-7.26 (m, 7H), 7.73 (d, 7= 8.1, 2H). LC-MS: 559.2 (M - H)', 99%,pure. Example 199. Preparation of Nα-isobutyl-Nα-(4-methylbenzenesulfonyl)-7Ve-(N,α- ethanolaminocarbonyl-L-phenylalanyl)-L-lysine
This compound was prepared as described for the preparation of Nα-isobutyl-Nα-(4- memylbenzenesulfonyl)-N€-[N'α-(4-memoxya
196) using ethanolamine instead of 4-methoxyaniline. The crude material was purified by preparative HPLC to give 4.1 mg, 5% of the desired material.
Η ΝMR (CDC13): δ 0.79 (d, 7= 6.3, 3H), 0.82 (d, 7= 6.3, 3H), 1.08-1.11 (m, 2H), 1.23-1.31 (m, 2H), 1.45-1.52 (m, IH), 1.92-1.97 (m, IH), 2.39 (s, 3H), 2.84-3.29 (m, 8H), 3.45-3.64 (m, 2H), 4.11 (br s, IH), 4.48 (br s, IH), 7.09-7.26 (m, 7H), 7.73 (d, 7= 8.1, 2H). LC-MS: 589.7 (M - H)\ 99% pure.
Example 200. Preparation of Nα-benzenesulfonyl-Nα-isobutyl-Ne-[N'α-(4- methyIbenzenesulfonyl)-L-phenylalanyl]-L-lysine
I Step A. Preparation of Nα-benzenesulfonyl-Nα-isobutyl-L-α-amino-e-caprolactam
!
I
Nα-isobuty'1-L-α-amino-e-caprolactam (example 1, step C) (4.0 g, 21.3 mmol, free base) was dissolved in DCM ( 100 mL) and treated with diisopropylethylamine (4.0 mL) followed by freshly recrystaUized benzenesulfonyl chloride (3.5 g, 21 mmol). The mixture was stirred overnight (TLC shows the reaction to be complete after 2 h). The solution was extracted with IN HCl and the organic layer was dried and evaporated to give 5.5 g (80%) of pure product. This compound was used without further purification in the next step.
Step B. Preparation of Nα-benzenesulfonyl-Nα-isobutyl-L-lysine
A mixture of Nα-benzenesulfonyl-Nα-isobutyl-L-α-amino-e-caprolactam (5.0 g, 15 mmol) and 6Ν HCl (50 mL) was refluxed for 6 h until all solids had disappeared. Afterwards, the solution was evaporated and the resulting solid was triturated with THF to give 5.2 g, 96% of the desired material. LC-MS: 346 (M + H)+, 99% pure.
Step C. Preparation of Nα-benzenesulfonyl-Nα-isobutyl-Ne-[N,α-(4-methylbenzenesulfonyl)-L- phenylalanyl]-L-lysine
A suspension ofNα-benzenesulfonyl-Nα-isobutyl-L-lysine (150 mg, 0.5 mmol), in THF (10 mL) was treated with a IN ΝaOH (3.0 mL) to pH 10. A solution of commercially available Nα-(4- methylbenzenesulfonyl)-L-phenylalanine acid chloride (187 mg, 0.5 mmol), in dry THF (10 mL) was added to the suspension and stirred for 4 h. Afterwards, water (2 mL) was added resulting in a clear solution. Then, EtOAc (30 mL) was added and the organic phase was washed with IN HCl. The organic phase was removed. Evaporation of the solvent gave a crude product which was purified by preparative HPLC to yield 19 mg (6%) of the title compound.
'HΝMR CCDCy: δ 0.76 (d, 7= 6.3, 3H), 0.80 (d, 7= 6.3, 3H), 1.10-1.20 (m, 2H), 1.26-133 (m, 2H), 1.70-1.74 (m,!lH), 1.89-1.93 (m, 2H), 2.38 (s, 3H), 279-2.90 (m, 2H), 3.85 (t, 7= 5.9, IH), 4.29 (t, 7= 6.9,; IH), 6.90 (d, 7 = 6.2, 2H), 7.08-7.29 (m, 6H), 7.35 (t, 7= 6.2, 2H), 7.44 (d, 7= 8.1, 2H), 773 (d57= 8.1, 2H). LC-MS: 642.8 (M - H)", 99% pure.
Example 201. Preparation ofNα-(4-aminobenzenesulfonyl)-Nα-isobutyl-Ne-[N'α-(4- methylbenzenesulfonyl)-L-phenylalanyI]-L-lysine
A suspension of Nα-(4-aminobenzenesulfonyI)-Nα-isobutyl-L-lysine potassium salt (190 mg, 0.5 mmol, example 166, step B) in THF (10 mL) was treated with a solution of Nα-(4- methylbenzenesulfonyl)-L-phenylalanine acid chloride (187 mg, 0.5 mmol) in dry THF (10 mL). The suspension was stirred for 4 h. Afterwards, water (2 mL) was added resulting in a clear solution. Then, EtOAc (30 mL) was added and the organic phase was washed with IN HCl. The organic phase was removed. Evaporation of the solvent gave a crude product which was purified by preparative HPLC to yield 15 mg (4.6%) of the title compound. (
]H ΝMR (CDC13): δ 0.74 (d, 7= 6.3, 3H), 0.80 (d, 7= 6.3, 3H), 1.00-1.11 (m, 4H), 1.23-1.25 (m, IH), 1.70-1.74 (m, IH), 1.89-1.93 (m, IH), 2.32 (s, 3H), 2.65 (m, 2H), 2.84-2.95 (ABX, 7 = 10.1, 7.1, 2H), 3.88 (t, 7= 6.0, IH), 4.11 (t, 7= 6.9, IH), 6.84 (d, 7= 6.6, IH), 7.02-7.21 (m, 7H), 7.24 (d, 7= 8.0, IH), 7.73 (d, 7= 8.1, 2H). LC-MS: 657.9 (M - H)\ 99% pure.
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Figure imgf000179_0001
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Figure imgf000179_0002
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Claims

We claim:
1. A compound selected from the group consisting of a compound of formula I ■
Figure imgf000180_0001
I
a compound of formula II.
Figure imgf000180_0002
and when the compound of formula I and II comprises an amino group pharmaceutically acceptable ammonium salts thereof,
wherein n is 3 or 4
wherein Y is O, S or N-CN
wherein Cx is selected from the group consisting of -COOM., COORg, -CHO, -CH2OR7, - CH2OCOR8, -CONHRg and -CONR]0RU, wherein M is an alkali metal or alkaline earth metal,
wherein R, is selected from the group consisting of a straight or branched alkyl group of 1 to 6 carbon atoms, a cycloalkylalkyl group having 3 to 6 carbon atoms in the cycloalkyl part thereof and 1 to 3 carbon atoms in the alkyl part thereof,
wherein R2 is selected from the group consisting of a benzenesulfonyl group of formula III,
Figure imgf000181_0001
III
and a thiophenesulfonyl group of formula IN,
Figure imgf000181_0002
IV
wherein R3 is selected from the group consisting of H, a straight or branched alkyl group of 1 to 6 carbon- atoms, a phenyl or a benzyl group .
wherein R4 is selected from the group consisting of H, a group of formula formula Ilia
Figure imgf000182_0001
πia
a group of formula INa
Figure imgf000182_0002
IVa
C6Hπ-, C5H10Ν-CH2CH2-, OC4H8N-CH2CH2-, C6H5CH2CH2-, 2,3-(CH3O)2C6H3CH2-, C6H5-, 2- C5H4N, 3-C5H4N, 4-C5H4N, 3-quinolyl, C6H5CS-, 2-naphthyl-SO2- and a group of formula R4C- CO-, R4C being selected from the group consisting of a straight or branched alkyl group of 1 to 6 carbon atoms, a cycloalkylalkyl group having 3 to 6 carbon atoms in the cycloalkyl part thereof and 1 to 3 carbon atoms in the alkyl part thereof, CF3, 1-pyrrolidinyl, 4-morpholinyl. tetrahydro- 3-furanyloxy, 4-CH3OC6H4NH-5 CH3NH-, HOCH2CH2NH-, 9-fluorenyl-CH2O-, tert-butylO-, /so- butylO-, C6H5CH2O-, CH3O-, unsubstituted C6H5-, C6H5- substituted by one or more members selected from the group consisting of F, Cl, Br, I, -CF3, -NO2, -NR-0R, ,, -NHCORi0s -OR10, -SR!0, -COOR10, -COR10 and -CH2OH, unsubstituted' C6H5CH2-, C6H5CH2- substituted by one or more members selected from the group consisting of F, Cl, Br, I, -CF3, -NO2, -NR.0Rn, -NHCOR-0. - OR10, -SR10, -COORI0, -COR10 and -CH2OH, unsubstituted C6H5CH2CH2-5 and C6H5CH2CH2- substituted by one or more members selected from the group consisting of F, Cl, Br, I, -CF3, - NO2, -NR-oR„, -NHCOR10, -OR10, -SR105 -COOR10, -COR10 and -CH2OH,
wherein R5 is selected from the group consisting of H, a straight or branched alkyl group of 1 to 8 carbon atoms, HOCH2-, C6H5CH2OCH2-3 benzyl-OCH(CH3), HO2CCH2-5 HO2CCH2CH2-, NC-CH2-, H2NC(O)CH2-, H2NC(O)CH2CH2-, 4-CH3C6H4CH2SCH2-5 CH3SCH2CH2-( H2NCH2CH2CH2CH2-, C6H5-, C6H5CH2-, C6H5CH(OH)-, C6H5CH(CN)-5 C6F5CH2-, 4-(9-fluorenylmethoxycarbonyl)-NHCH2- C6H4CH2-5 C5H4N-2-CH2-, C5H4N-3-CH2-, C5H4N-4-CH2-, 2-thiophene-CH2-_ indole-3-CH2-, 2- benzothiophene-CH2-, Nτ-benzyl-imidazole-4-CH2-, imidazole-4-CH2-, thiazole-4.-CH2- and benzyl substituted by a group selected from the group consisting of H, a straight or branched alkyl group of 1 to 4 carbon atoms, F, Cl, Br, I, -CF3, -ΝO2, -NR10Rn, -NHCOR10s -OR10, -OCH2C6H5, -SR10, -COOR-o, -COR10 and -CH2OH,
wher
Figure imgf000183_0001
wherein Met is a methylene linked to the α' nitrogen
wherein _j is selected from the group consisting of H, a straight or branched alkyl group of 1 to 4 carbon atoms and glycyl
wherein R7 is selected from the group consisting of H, a straight or branched alkyl group of 1 to 4 carbon atoms
wherein R8 is selected from the group consisting of a straight or branched alkyl group of 1 to 6 carbon atoms, a cycloalkyl group having 3 to 6 carbon atoms, a cycloalkylalkyl group having 3 to 6 carbon atoms in the cycloalkyl part and 1 to 3 carbon atoms in the alkyl part thereof,
wherein Rg is selected from the group consisting of H, a straight or branched alkyl group of 1 to 4 carbon atoms, -OH, -NH2 and -CH2CH2OH • wherein R10 and Rn are independently selected from the group consisting of H, a straight or branched alkyl group of 1 to 4 carbon atoms
wherein m is 0 or 1
wherein o is 0 or 1
wherein R12 and R13 are independently selected from the group consisting of H, a straight or branched alkyl group of 1 to 4 carbon atoms, F, Cl, Br, I, -CF3. -NO2, -NR-0Rn, -NHCOR10, - OR,0, -OCH2C6H5, -SR-o, -COOR10. -COR10 and -CH2OH, R10 and Rn being as defined above.
2. A compound of formula IA
Figure imgf000184_0001
IA
and when the compound of formula IA comprises an amino group pharmaceutically acceptable ammonium salts thereof,
wherein n is 3 or 4
• ' ■ ■ I . • • wherein Y is 6, S or N-CN
wherein Cx is selected from the group consisting of -COOM, COOR^ -CHO, -CH2OR7- - CH2OCOR8, -CONHRg and -CONR,0R-,5 wherein M is an alkali metal or alkaline earth metal, wherein R- is selected from the group consisting of a straight or branched alkyl group of 1 to 6 carbon atoms, a cycloalkylalkyl group having 3 to 6 carbon atoms in the cycloalkyl part thereof and 1 to 3 carbon atoms in the alkyl part thereof,
wherein R2 is selected from the group consisting of a benzenesulfonyl group of formula III,
Figure imgf000185_0001
III
I - i
and a thiophenesulfonyl group of formula IV,
Figure imgf000185_0002
IV
wherein R3 is selected from the group consisting of H, a straight or branched alkyl group of 1 to 6 carbon atoms, a phenyl or a benzyl group ;
wherein R4 is selected from the group consisting of H. C6Hπ-, CjH10N-CH2CH2-, OC4H8N- CH2CH2-, C6H5CH2CH2-3 2,3-(CH3O)2C6H3CH2-, C6H5-5 2-C5H4N, 3-C5H4N, 4-C5H4N, 3- quinolyl, CH3CO-3 CF3CO, C6H5CO-, C6H5CS-3 4-CH3OC6H4CH2CO-3 C6H5CH2CH2CO-5 iso- butyl-CO-5 z5O-propyl-CO-, tert-butyl-CO-, tert-butyl-CH2CO-, l-pyrrolidine-CO-^4-morpholine- CO-, carbotetrahydro-3-furanyloxy, 4-CH3OC6H4NHCO-, CH3NHCO-5 HOCH2CH2NHCO-- 9- fluorenyl ethoxycarbonyl, tert-butylO-CO-, z'so-butylO-CO-, C6H5CH2O-CO-, CH3O-CO-5 C6H5SO2-, 4-CH3C6H4SO2-5 4-CF3C6H4SO2-, 4-NO2C6H4SO2-3 4-NH2C6H4SO2-3 ' 4- AcNHC6H4SO2-, 4-FC6H4SO2-34-ClC6H4SO2-, 4-BrC6H4SO2-, 4-CH3OC6H4SO2-32-thiophene- SO2- and 2-naphthyl-SO2-
wherein R5 is selected from the group consisting of H, CH3-3 CH3CH2CH2-,CH3CH2CH2CH2-, wø-C4H9-3 C6HnCH2-, HOCH2-3 C6H5CH2OCH2-, benzyl-OCH(CH3), HO2CCH2-3 HO2CCH2CH2- , NC-CH2-3 H2NC(O)CH2-, H2NC(O)CH2CH2-3 4-CH3C6H4CH2SCH2-, C ^SCH^H^ 2HNCH2CH2CH2CH2-3 C6H5-3 C6H3CH2-3 C6F5CH2-34-tert-butyl-C6H4CH2-34-HOC6H4CH2-, 4- benzyl-O-C6H4CH2-5 4-NO2C6H4CH2-, 4-(9-fluorenylmethoxycarbonyl)NHCH2-C6H4CH2-, 2- FC6H4CH2-3 3-FC6H4CH2-3 4-FC6H4CH2-3 C5H4N-2-CH2-3 C5H4N-3-CH2-3 C5H4N-4-CH2-3 2- thiophene-CH2-> indole-3-CH2-3 2-benzothiophene-CH2-3 Nτ-benzyl-imidazole-4-CH2- and thiazole-4-CH2-
wherein R^ is selected from the group consisting of H, a straight or branched alkyl group of 1 to 4 carbon atoms and glycyl
wherein R7 is selected from the group consisting of H, a straight or branched alkyl group of 1 to 4 carbon atoms ; , ) ■
wherein R8 is selected from the group consisting of a straight or branched alkyl group of 1 to 6 carbon atoms, a cycloalkyl group having 3 to 6 carbon atoms, a cycloalkylalkyl group having 3 to 6 carbon atoms in the cycloalkyl part and 1 to 3 carbon atoms in the alkyl part thereof,
wherein g is selected from the group consisting of H, a straight or branched alkyl group of 1 to 4 carbon atoms, -OH, -ΝH2 and -CH2CH2OH
wherein R,0 and R, ■ are independently selected from the group consisting of H, a straight or branched alkyl group of 1 to 4 carbon atoms
wherein m is O or l
wherein o is 0 or 1 wherein R12 and R]3 are independently selected from the group consisting of H, a straight or branched alkyl group of 1 to 4 carbon atoms, F, Cl, Br, I, -CF3, -NO2, -NR-0Rn, -NHCOR10, - OR]0, -OCH2C6H5, -SR10, -COOR]0, -COR-0 and -CH2OH, R]0 and Rπ being as defined above.
3. A compound as defined in claim 2 wherein Cx is -CO2H, Y is O and n is 4.
4. A compound as defined in claim 3 wherem R- is t_obutyl.
5. A compound of formula II as defined in claim 1 wherein Cx is -CO2H, Y is O, n is 4 and wherein R^ represents a member selected from the group consisting of
Figure imgf000187_0001
wherein Met is a methylene linked to the α' nitrogen.
6. A compound as defined in claim 2 wherein Cx is -CO2H, Y is S and n is 4.
7. A compound of formula IA as defined in claim 3 wherein R- is selected from iso- utyl and cyclopentyl-CH2- and R2 is selected from 4-CH3C6H4SO2-, 4-NH2C6H4SO,-, 4-NO2C6H4SO2-, 2-thiophene-SO2-, 4-CH3OC6H4SO2- and C6H4SO2-.
8. A compound of formula IA as defined in claim 4 wherein R2 is selected from 4-CH3C6H4SO2-, 4-NH2C6H4SO2-, 4-NO2C6H4SO2-, 2-thiophene-SO2-, 4-CH3OC6H4SO2- and C6H4SO2-.
9. A compound as defined in claim 6 wherein R, is wo-butyl and R2 is 4-CH3C6H4SO2-.
10. A compound of formula and K, N
11. A co
Figure imgf000188_0001
and K, Na and Cs salts thereof.
12. A compound of formula
Figure imgf000188_0002
and K, Na and Cs salts thereof.
13. A compound of formula
Figure imgf000189_0001
pharmaceutically acceptable ammonium salts thereof and K3 Na and Cs salts thereof.
14. A compound of formula
Figure imgf000189_0002
pharmaceutically acceptable ammonium salts thereof and K, Na and Cs salts thereof.
15. A compound of formula
Figure imgf000189_0003
and K. Na and Cs salts thereof.
16. A compound of formula
Figure imgf000190_0001
and K, Na and Cs salts thereof.
17. A compound of formula
Figure imgf000190_0002
pharmaceutically acceptable ammonium salts thereof and K, Na and Cs salts thereof.
18. A compound of formula
Figure imgf000190_0003
pharmaceutically acceptable ammonium salts thereof and K, Na and Cs salts thereof.
19. A compound of formula
Figure imgf000191_0001
and K3 Na and Cs salts thereof.
20. A compound of formula
Figure imgf000191_0002
and K, Na and Cs salts thereof.
21. A compound of formula
Figure imgf000191_0003
and K3 Na and Cs salts thereof.
22. A compound of formula
Figure imgf000192_0001
and K, Na and.Cs salts thereof.
23. A compound of formula
Figure imgf000192_0002
and K, Na and Cs salts thereof.
24. A compound of formula
Figure imgf000192_0003
pharmaceutically acceptable ammonium salts thereof and K, Na and Cs salts thereof.
25. A compound of formula
Figure imgf000193_0001
26. A compound of formula
Figure imgf000193_0002
27. A compound of formula
Figure imgf000193_0003
28. A compound of formula
Figure imgf000194_0001
and K, Na and Cs salts thereof.
29. A compound of formula
Figure imgf000194_0002
and K. Na and Cs salts thereof.
30. A compound of formula
Figure imgf000194_0003
31. A compound of formula
Figure imgf000195_0001
pharmaceutically acceptable ammonium salts thereof and K, Na and Cs salts thereof.
A compound of formula
Figure imgf000195_0002
and K, Na and Cs salts thereof.
33. A compound of formula
Figure imgf000196_0001
and pharmaceutically acceptable ammonium- salts thereof.
54/ A compound of formula
Figure imgf000196_0002
and phamiaceuticaily acceptable ammonium salts thereof.
35. A compound of formula
Figure imgf000196_0003
and K, Na and Cs salts thereof.
36. A compound of formula
Figure imgf000197_0001
37. A compound of formula
Figure imgf000197_0002
and pharmaceutically acceptable ammonium salts thereof.
38. A compound of formula
Figure imgf000197_0003
and pharmaceutically acceptable ammonium salts thereof.
39. A compound of formula
Figure imgf000198_0001
40. A compound of formula
Figure imgf000198_0002
and phamiaceuticaily acceptable ammonium salts thereof.
41. A compound of formula
Figure imgf000198_0003
and pharmaceutically acceptable ammonium salts thereof.
42. A compound of formula
Figure imgf000199_0001
and pharmaceutically acceptable ammonium salts thereof.
43. A compound of formula
Figure imgf000199_0002
44. A compound of formula
Figure imgf000199_0003
45. A compound of fomiula
Figure imgf000199_0004
and phamiaceuticaily acceptable ammonium salts thereof.
46. A compound of formula
Figure imgf000200_0001
and pharmaceutically acceptable ammonium salts thereof.
47. A compound of fomiula
Figure imgf000200_0002
and phamiaceuticaily acceptable ammonium salts thereof.
48. A compound of formula
Figure imgf000200_0003
49. A compound of formula
Figure imgf000201_0001
50. A pharmaceutical composition comprising a pharmaceutically acceptable carrier and a pharmaceutically effective amount of at least one compound of formula IA as defined in claim 2.
51. A compound, selected from the group consisting of a compound of formula la
Figure imgf000201_0002
and when the compound of formula la comprises an amino group pharmaceutically acceptable ammonium salts thereof,
wherein Cx is selected from the group consisting of -COOM and -CH2OH, M being an alkali metal or alkaline earth metal,
wherein R]A is selected from the group consisting of a straight or branched alkyl group of 1 to 6 carbon atoms, a cycloalkylalkyl group having 3 to 6 carbon atoms in the cycloalkyl part thereof and 1 to 3 carbon atoms in the alkyl part thereof, wherein R2A and R4A are independently selected from the group consisting of H, a straight or branched alkyl group of 1 to 4 carbon atoms, F, Cl, Br, I, -CF3, -NO2, -NR]0Rn, -NHCOR10, - ORI0, -OCH2C6H5, -SRI0, -COOR,o, -COR]0 and -CH2OH,
wherein R5A is selected from the group consisting of H, a straight or branched alkyl group of 1 to 8 carbon atoms, HOCH2-, C6H5CH2OCH2-, benzyl-OCH(CH3), HO2CCH2-, HO2CCH2CH2-, NC-CH2-, H2NC(O)CH2~, H2NC(O)CH2CH2-, 4-CH3C6H4CH2SCH2-, CH3SCH2CH2- H2NCH2CH2CH2CH2-, C6H5-, C6H5CH2-, C6HsCH(OH)-, C6H5CH(CN)-, C6F5CH2-,4-(9- fluorenylmethoxycarbonyl)-NHCH2-C6H4CH2-, C5H4N-2-CH2-, C5H4N-3-CH2-, C5H4N-4-CH2-, 2-thiophene-CH2-ι indole-3-CH2-, 2-benzothiophene-CH2-, Nτ-benzyl-imidazole-4-CH2-, imidazole-4-CH2-, thlazole-4-CH2- and benzyl substituted by a group selected from the group consisting of H, a straight or branched alkyl group of 1 to 4 carbon atoms, F, Cl, Br, I, -CF3, - ΝO2, -NR10Rn. -NHCOR103 -OR10, -OCH2C6H5, -SR10, -COOR10, -COR10 and -CH2OH,
and wherein R)0 and R-- are independently selected from the group consisting of H, a straight or branched alkyl group of 1 to 4 carbon atoms.
52. A compound selected from the group consisting of a compound of formula lb
Figure imgf000202_0001
and when the compound of formula lb comprises an amino group pharmaceutically acceptable ammonium salts thereof, » .
wherein Cx is selected from the group consisting of -COOM, and -CH2OH, M being an alkali metal or alkaline earth metal, wherein R1A is selected from the group consisting of a straight or branched alkyl group of 1 to 6 carbon atoms, a cycloalkylalkyl group having 3 to 6 carbon atoms in the cycloalkyl part thereof and 1 to 3 carbon atoms in the alkyl part thereof,
wherein R2A and R4B are independently selected from the group consisting of H, a straight or branched alkyl group of 1 to 4 carbon atoms, F, Cl, Br, I, -CF3, -NO2, -NR-0R-„ -NHCOR10, - OR10, -OCH2C6H5, -SR10, -COOR10, -COR10 and -CH2OH,
wherein R5A is selected from the group consisting of H, a straight or branched alkyl group of 1 to 8 carbon atoms, HOCH2-, C6H5CH2OCH2-, benzyl-OCH(CH3), HO2CCH2-, HO2CCH2CH2-, NC-CH2-, H2NC(O)CH2-, H2NC(O)CH2CH2-, 4-CH3C6H4CH2SCH2-, CH3SCH2CH2-, H2NCH2CH2CH2CH2-, C6H5-, C6H5CH2-, C6H5CH(OH)-, C6H5CH(CN)-, C6FSCH2-, 4-(9- fluorenylmethoxycarbonyl)-NHCH2-C6H4CH2-, C5H4N-2-CH2-, CsH4N-3-CH2-, C5H4N-4-CH2-, 2-thiophene-CH2-> indole-3-CH2-, 2-benzothiophene-CH2-s Nτ-benzyl-imidazole-4-CH2-, imidazole-4-CH2-, thiazole-4-CH2- and benzyl substituted by a group selected from the group consisting of H, a straight or branched alkyl group of 1 to 4 carbon atoms, F, Cl, Br, I, -CF3, - ΝO2, -NR]0R„, -NHCOR-o, -OR10, -OCH2C6H5, -SR]0, -COOR10, -COR10 and -CH2OH,
and wherein R10 and R-- are independently selected from the group consisting of H, a straight or branched alkyl group of 1 to 4 carbon atoms.
53. A compound of selected, from the group consisting of a compound of formula Ic
Figure imgf000203_0001
and when the compound of formula Ic comprises an amino group pharmaceutically acceptable ammonium salts thereof,
wherein Cx is selected from the group consisting of -COOM and -CH2OH, M being an alkali metal or alkaline earth metal,
wherein R,A is selected from the group consisting of a straight or branched alkyl group of 1 to 6 carbon atoms, a cycloalkylalkyl group having 3 to 6 carbon atoms in the cycloalkyl part thereof and 1 to 3 carbon atoms in the alkyl part thereof,
wherein R2B and R4B are independently selected from the group consisting of H, a straight or branched alkyl group of 1 to 4 carbon atoms, F, Cl, Br, I, -CF3, -NO2, -NR.0Rπ, -NHCOR-0, - OR10, -OCH2C6H5, -SR10, -COOR10, -COR10 and -CH2OH,
wherein RJA is selected from the group consisting of H, a straight or branched alkyl group of 1 to 8 carbon atoms, HOCH2-, C6H5CH2OCH2-, benzyl-OCH(CH3), HO2CCH2-, HO2CCH2CH2-, NC-CH,-, H2NC(O)CH2-, FI2NC(O)CH2CH2-, 4-CH3C6H4CH2SCH2-, CH^CH^H^ H2NCH2CH2CH2CH2-, C6H5-, C6H5CH2-, C6H5CH(OH)-, C6H5CH(CN)-, C6F5CH2-, 4-(9- fluorenylmethoxycarbonyl)-NHCH2-C6H4CH2~, C5H4N-2-CH2-, C5H4N-3-CH2-, C5H4N-4-CH2-, 2-thiophene-CH2-j indole-3-CH2-, 2-benzothiophene-CH2-, Nτ-benzyl-imidazole-4-CH2-, imidazole-4-CH2-, thiazole-4-CH2- and benzyl substituted by a group selected from the group consisting of H, a straight or branched alkyl group of 1 to 4 carbon atoms, F, Cl, Br, I, -CF3, - ΝO2, -NR10RU :, -NHCOR-o, -OR10, -OCH2C6H5, -SR10, -COOR10, -COR10 and -CH2OH,
and wherein RI0 and R, - are independently selected from the group consisting of H, a straight or branched alkyl group of 1 to 4 carbon atoms.
54. A compound of selected from the group consisting of a compound of formula Id
Figure imgf000205_0001
and when the compound of formula Id comprises an amino group pharmaceutically acceptable ammonium salts thereof, ,
wherein Cx is selected .from the group consisting of -COOM and -CH2OH, M being an alkali metal or alkaline earth metal,
wherein R1A is selected from the group consisting of a straight or branched alkyl group of 1 to 6 carbon atoms, a cycloalkylalkyl group having 3 to 6 carbon atoms in the cycloalkyl part thereof and 1 to 3 carbon atoms in the alkyl part thereof,
wherem R4A is selected from the group consisting of H, a straight or branched alkyl group of 1 to 4 carbon atoms, F, Cl, Br, I, -CF3, -NO2, -NR-0R-„ -NHCOR10, -OR10, -OCH2C6H5, -SR10, - COOR10, -COR10 and -CH2OH,
wherein R4C is selected from the group consisting of a straight or branched alkyl group of 1 to 6 carbon .atoms, a cycloalkylalkyl group having 3 to 6 carbon atoms in the cycloalkyl part thereof and 1 o 3 carbon atoms in the alkyl part thereof, CF3, pyrrolidine, 4-morpholine, tetrahydro-3- furanyloxy, 4-CH3OC6H4NH-, CH3NH-, HOCH2CH2NH-, 9-fluorenyl-CH2O-, tert-butylO-, zso- butylO-, C6H5CH2O-, CH3O-, unsubstituted C6H5-, C6H5- substituted by one or more members selected from the group consisting of F, Cl, Br, I, -CF3, -NO2, -NR!0Rn, -NHCOR10, -OR10, -SR10, -COOR-o, -COR10 and -CH2OH, unsubstituted C6H5CH2-, C6H5CH2- substituted by one or more members selected from the group consisting of F, Cl, Br, I, -CF3, -NO2, -NR-0Rn, -NHCOR10, - OR10, -SR10, -COOR]0, -COR10 and -CH2OH, unsubstituted C6H5CH2CH2-, and C6H5CH2CH2- substituted by one or more members selected from the group consisting of F, Cl, Br, I, -CF3, - NO2, -NR-oRn, -NHCOR]0, -OR10, -SR,0, -COOR10, -COR10 and -CH2OH,
wherein R5A is selected from the group consisting of H, a straight or branched alkyl group of 1 to 8 carbon atoms, HOCH2-, C6H5CH2OCH2-, benzyl-OCH(CH3), HO2CCH2-, HO2CCH2CH2-, • NC-CH2-, H2NC(O)CH2-, H2NC(O)CH2CH2-, 4-CH3C6H4CH2SCH2-, CH^CH^H^ H2NCH2CH2CH2CH2-, C6H5~, C6H5CH2-, C6H5CH(OH)-, C6H5CH(CN)-, C6F5CH2-, 4-(9- fluorenylmethoxycarbonyl)-NHCH2-C6H4CH2-, C5H4N-2-CH2-, C5H4N-3-CH2-, C5H4N-4-CH2-, 2-thiophene-CH2-ι indole-3-CH2-, 2-benzothiophene-CH2-, Nτ-benzyl-imidazole-4-CH2- , imidazole-4-CH2-3 thiazole-4-CH2-3 and benzyl substituted by a group selected 'from the group consisting of H, a straight or branched alkyl group of 1 to 4 carbon atoms, F, Cl, Br, I, -CF3, - ΝO2, -NR]0R.., -NHCOR-o, -OR10, -OCH2C6H5, -SR10, -COOR10, -COR10 and -CH2OH,
and wherein R10 and Rn are independently selected from the group consisting of H, a straight or branched alkyl group of 1 to 4 carbon atoms. -
55. A compound as defined in claim 1 wherein R4 is selected from the group consisting of C6H5SO2-, 4-CH3C6H4SO2-, 4-CF3C6H4SO2-, 4-NO2C6H4SO2-, 4-NH2C6H4SO2-, 4- AcNHC6H4SO2-, 4-FC6H4SO2-, 4-ClC6H4SO2-, 4-BrC6H4SO2-, 4-CH3OC6H4SO2-, and 2- thiophene-SO2-, CH3CO-, CF3CO-, C6H5CO-, 4-CH3OC6H4CH2CO-, C6H5CH2CH2CO-, iso- butyl-CO-, /.SO-propyl-CO-, tert-butyl-CO-, tert-butyl-CH2CO-, pyrrolidine-CO-, 4-morpholine- CO-, carbotetrahydro-3-furanyloxy, 4-CH3OC6H4NHCO-, CH3NHCO-, HOCH2CH2NHCO-, 9- fluorenylmethoxycarbonyl, tert-butylO-CO-, wo-butylO-CO-, C6H5CH2O-CO-, and CH3O-CO- and wherein R5 is selected from the group consisting of H, CH3-, CH3CH2CH2-,CH3CH2CH2CH2-, is o-C4H9-, C6Hn CH2-, HOCH2-, C6H5CH2OCH2-, benzyl-OCH(CH3), HO2CCH2-, HO2CCH2CH2- , NC-CH2-3 H2NC(O)CH2-, H2NC(O)CH2CH2-5 4-CH3C6H4CH2SCH2-, CH3SCH2CH2-( H2NCH2CH2CH2CH2-, C6H5-, C6H5CH2-, C6H5CH(OH)-, C6H5CH(CN)-, C6F5CH2-, 4-tert-butyl- C6H4CH2-, 4-HOC6H4CH2-, 4-benzyl-O-C6H4CH2-, . -NO2C6H4CH2-, 4-(9- fluorenylmethoxycarbonyl)NHCH2-C6H4CH2-, 2-FC6H4CH2-, 3-FC6H4CH2-, 4-FC6H4CH2-, C5H4N-2-CH2-, C5H4N-3-CH2-, C5H4N-4-CH2-, 2-thiophene-CH2-ι indole-3-CH2-, 2- benzothiophene-CH2-, Nτ-benzyl-imidazole-4-CH2- , imidazole-4-CH2-, and thiazole-4-CH2-. .
56. The trifluoroacetic ammonium salt of a compound of formula IA as defined in claim 2 wherein R3 and R4 are H.
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US6506786B2 (en) 2003-01-14
US6656965B2 (en) 2003-12-02
US20020151546A1 (en) 2002-10-17
US6610689B2 (en) 2003-08-26
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US6677367B2 (en) 2004-01-13
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CA2440931C (en) 2009-09-08
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US20030119803A1 (en) 2003-06-26
US20030195159A1 (en) 2003-10-16

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